Journal of the Korean Physical Society (2022) 80:817–851 Online ISSN 1976-8524

https://doi.org/10.1007/s40042-022-00442-w Print ISSN 0374-4884

ORIGINAL PAPER - FLUIDS, PLASMA AND PHENOMENOLOGY

Plasma bioscience for medicine, agriculture and hygiene applications

Eun Ha Choi1 · Nagendra Kumar Kaushik1 · Young June Hong1 · Jun Sup Lim1 · Jin Sung Choi1 · Ihn Han1

Received: 25 August 2021 / Accepted: 19 October 2021 / Published online: 4 March 2022
© The Korean Physical Society 2022

Abstract
Nonthermal biocompatible plasma (NBP) sources operating in atmospheric pressure environments and their characteristics
can be used for plasma bioscience, medicine, and hygiene applications, especially for COVID-19 and citizen. This review
surveyed the various NBP sources, including a plasma jet, micro-DBD (dielectric barrier discharge) and nanosecond dis-
charged plasma. The electron temperatures and the plasma densities, which are produced using dielectric barrier discharged
electrode systems, can be characterized as 0.7 ~ 1.8 eV and (3–5) × ­1014–15 ­cm−3, respectively. Herein, we introduce a general
schematic view of the plasma ultraviolet photolysis of water molecules for reactive oxygen and nitrogen species (RONS)
generation inside biological cells or living tissues, which would be synergistically important with RONS diffusive propa-
gation into cells or tissues. Of the RONS, the hydroxyl radical [OH] and hydrogen peroxide ­H2O2 species would mainly
result in apoptotic cell death with other RONS in plasma bioscience and medicines. The diseased biological protein, cancer,
and mutated cells could be treated by using a NBP or plasma activated water (PAW) resulting in their apoptosis for a new
paradigm of plasma medicine.

Keywords Nonthermal biocompatible plasma (NBP) · Plasma bioscience and medicine · Hygiene applications · Virus
inactivation · Cancer treatment · Plasma jet · Micro-DBD · Nanosecond discharged plasma · Plasma ultraviolet photolysis ·
Plasma activated water (PAW)

1 Introduction charged ions, neutral molecules in ground states and reactive

A plasma is a group of electrically charged particles along
with visible light, infrared, ultraviolet rays, neutral gases,
excited reactive species, and some heat. It is the so-called
fourth state of matters, the other three being the solid, liq-
uid, and gaseous states. A plasma can be produced by using
either a direct current (DC) or an alternating current (AC)
electric discharge in a gas between powered and grounded
electrodes [1–8]. The electric discharges produce electrons,
ions, light, heat, and reactive neutral gases in high-energy
excited states throughout the discharge processes. This mat-
ter is the basic substance for the creation of the universe,
which is referred to in physics, as well as the source of life-
forming substances in medical life science. The constituent
gas of the plasma is negatively charged electrons, positively
* Eun Ha Choi
ehchoi@kw.ac.kr
1
Department of Electrical and Biological Physics, Plasma
Bioscience Research Center and Applied Plasma Medicine
Center, Kwangwoon University, Seoul 01897, Korea
oxygen and nitrogen species (RONS) with high chemical
reactivity with their neighboring molecules [7–11].
The meaning of the plasma can be found more accurately
by looking for a Sanskrit प्लाज्मा (plasma) rather than a
Greek (πλασμα) word. The Sanskrit word “Pla or pra” means
very basic, primitive, very high, “z, s, or sura” means life or
water, and “ma” means a collection of energy, i.e., matter
[12]. Hence a plasma can be interpreted as a “fundamental
and life substance or material in an universe with a highly
accumulated state of energy”. In medicine and biology,
because “blood” and “proplasm” have been thought of as
the primitive and essential material of life [13–15], they are
called as “bio-plasma” and “plasma”, respectively. Around
100 years ago, Irvine Langmuir named this kind of electri-
cally discharged gas stream species as “plasma” after these
properties which are similar to those of biological fluids.
In 1953, Stanley Miller performed discharge experiments
under primitive atmospheric environments consisting of
ammonia, methane, water vapor, and hydrogen gases and
observed and watched the formation of amino acid such as
alanine, glycine, and others [16]. These two or three amino

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acids are constituent molecules that can be combined to form
a protein, which is the basic substance of life. Furthermore,
life substances are known to be born through their polym-
erization. Meanwhile, an April 25, 1953, biologist Watson
and physicist Crick reported that deoxyribose nucleic acid
(DNA) is a genetic factor consisting of a double helix struc-
ture, which can give a deeper understanding of the molecular
biology in fundamental life [17]. In modern medicine, all
kinds of diseases, such as skin, dementia, cancers and den-
tals, occur due to the sudden mutation and oxidation of these
cells. Proteins could have been generated by the synthesis
of amino acids in the discharge state of the earth’s primi-
tive atmospheric environments. Based on these facts, we can
treat the degenerate neural Alzheimer's and Parkinson's dis-
eases, atopy, wounds and cancer. A hypothesis that plasma
treatment is fundamentally possible may be established and
realized for the next generation of plasma bioscience and
medicine [5, 18–22]. Furthermore, it can be used in agricul-
ture [23–34], oral health [35–43], and public health [44–49].
Low-temperature plasmas generated at ambient atmos-
pheric pressure generally can be taken to have a cocktail
form in which the RONS are mixed with electrons, ions,
other neutrals and ultraviolet (UV) light. In particular, this
type of nonthermal biocompatible plasma (NBP) is simply
referred to as a nonthermal atmospheric pressure plasma
(NAP) or a cold atmospheric pressure plasma (CAP) [5, 6,
11, 19, 50–65]. Here, we may call them simply as NBP. This
kind of NBP has a slight thermal effect, as well as electrons
and ions, reactive oxygen and nitrogen gases, ultraviolet and
visible light. Figure 1a shows that the reactive neutral mol-
ecules with highly excited-energy in a NBP have strong reac-
tivity with other molecules in prokaryotic and eukaryotic
cells, as well as cell membrane and deoxyribose nucleic acid
(DNA), by taking electrons from surrounding biomolecules
or atom [66]. At this time, the atoms or molecules losing
electrons are said to have been oxidized.
­1012 to ~ ­1016 ­cm−3 and electron temperatures from ~ 0.8 to
Fig. 1  Plasma bioscience and medicine with a nonthermal biocom-
patible plasma (NBP) [14]
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Among the reactive species, the hydroxyl radical [OH],
atomic oxygen [O], hydrogen peroxide [­ H2O2], singlet oxy-
gen [↑O2↓], nitric oxide [NO], superoxide anion [­ O2*−],
ozone ­[O3], peroxynitrite [­ ONOO−], nitrite [­ NO2−], nitrate
­[NO3−], and excited nitrogen molecules [­ N2*] play crucial
roles in interactions with cells or microbials, as do their
positive ions and electrons [5, 53, 54, 60]. Also, electromag-
netic waves, such as ultraviolet, visible and infrared rays, are
emitted from the excited gases of helium (He), argon (Ar),
nitrogen ­(N2), air, and their mixtures during the discharges
in plasma. Interactions of these plasma generated RONS
and electromagnetic waves with biological cells, tissues and
microbials, such as bacteria, fungi, and viruses, can be used
for wound treatment for tissue regeneration, selective cancer
apoptosis, microbial biofilm removal, and public health, as
shown in Fig. 1. Figure 1 shows selective ROS targeting can-
cer cells yielding to apoptosis, while normal cells experience
proliferation, according to their exposed energy to plasma.
The normal levels of ROS contents in normal and cancer
cells are low and high, respectively [67]. If a plasma is used
to treat both cells, then the ROS levels in those cells are
increased; however, they are lower in normal cells and quite
a bit higher than cytotoxicity threshold for inducing apopto-
sis in cancer cells, respectively [67]. Hence, the cancer cells
selectively can be caused to undergo apoptosis due to a NBP.
Also, plasma medical devices, along with their safety
standards, can be developed through research on biological
interactions with plasma. Also, necessary data for plasma
medical device development can be acquired through
direct and indirect treatment of abnormal cell populations.
Moreover, plasma treated water (PTW) [68–74], which is
made by bombarding water with plasma, resulting in long-
lived residual radicals and cells, can be used to interact
with living tissues for many purposes.
The purpose of this review is to provide current stud-
ies and recent results of plasma bioscience and medicine
based on plasma physics, chemistry, biology, and agricul-
ture. Also, we review typical NBP sources, their plasma
diagnostics, and their applications in cancer treatment,
agriculture, and microbial treatment for bacteria removal
and virus inactivation. Most NBP plasma sources are
based on dielectric barrier discharged (DBD) plasmas,
in which both or one of the electrodes is blocked by a
dielectric material between them. There are two types
of DBD plasmas: a pencil-type plasma jet and a surface
discharged or facing discharged DBD plasma. These are
frequently used with driving frequencies from less than
1 kHz up to microwave frequency ~ GHz. These NBP plas-
mas can be characterized by their electron densities from
3 eV [8, 14]. The plasma gas temperature in NBP plumes
is very important for patients because heat at a temperature
above 45 °C is not allowed by regulation.
Plasma bioscience for medicine, agriculture and hygiene applications 819
2 Nonthermal biocompatible plasma (NBP)
generator operating at atmospheric
pressure
2.1 Nonthermal biocompatible plasma (NBP)
generator
Many dielectric barrier discharge (DBD) plasma sources
[57] for plasma medicines have been developed since E.
W. von Siemens first studied them in 1857. These are
known to be AC silent discharges with nonequilibrium
atmospheric pressure plasma characteristics. DBD plasma
sources can be manufactured using two types of geometri-
cal configurations: a typical atmospheric-pressure nitrogen
(or air) plasma jet based on DBD geometry as shown in
Fig. 2a, and coplanar DBD (C-DBD) surface plasma types
as shown in Fig. 2b. The plasma jet, which is currently
very popular worldwide due to its ease of manufacture,
and the C-DBD plasma whose electrode is based on a pat-
ent from plasma display panel technology [75, 76], can be
used by the PBRC (Plasma Bioscience Research Center) to
lead the development of health medicine, agriculture and
fisheries, beauty, and environmental improvement for air,
water and soil. The plasma jet system consists of a high
voltage power supply and electrodes covered with dielec-
trics. A plasma jet device is assembled with a syringe and
a glass or ceramic tube. A duty-cycled sinusoidal wave
with a frequency of 60 Hz–1 GHz can be used in a plasma
jet with a few kilovolts. A medical syringe or needle is
used as a powered electrode and as guiding tube for the gas
flow, and is made of stainless steel with an inner diameter
is 1.2–3.3 mm and a thickness is 0.1–0.3 mm, which is
tightly surrounded by a quartz tube whose outer diameter
is 7–10 mm. The glass or quartz tube covering the needle
also prevents electric shocks caused by high voltage across
Fig. 2  a Schematic of a typical atmospheric-pressure nitrogen (or air) plasma jet device with a 3 lpm nitrogen (or air) flow and the quartz dielec-
tric tube; and b coplanar dielectric barrier discharged (C-DBD) surface plasma [77]
a bare needle and plays the role of a nozzle for the plasma
plume. The outer grounded electrode is placed outside the
end of the glass tube and is made of stainless steel with
a central hole of 1 mm in diameter, through which the
plasma is ejected into the surrounding ambient air. The
discharge gap distance is adjusted to 2–3 mm between the
inner and outer electrode. Air and any other gases or their
mixtures can be used as the feeding gas. The electrical dis-
charge can be produced and sustained by using two elec-
trodes barriered by dielectric materials. These dielectrics
are commonly made of glass, bakelite, quartz, ceramics,
and polymers. Energetic electrons and ions are generated
first on the dielectric surface discharge path, followed by
volume discharges caused by ionization in the gases. Cells
or biological targets have been treated with plasma jets
for appropriate exposure times. The working temperature
of the plasma source is in the range of 26–36 °C during
plasma treatment [77].
Figure 2b shows a coplanar dielectric barrier discharged
(C-DBD) plasma operating at ambient atmospheric pressure
for biosciences, medicines and esthetics [77]. Biological tis-
sue or skin have been treated by using C-DBDs for 30 s and
a few minutes with operating voltages of 1–2.2 kV and dis-
charge current 1–2 mA. The C-DBD plasma device consists
of two parallel silver electrodes screen printed on a glass
or ceramic substrate. The coplanar two electrodes whose
thickness are about 3–5 um are separated by 100–200 um,
and tightly screen printed by using ­SiO2 paste with a thermal
treatment at 600 °C. C-DBD plasma can be manufactured
with diameter from 30 to 300 mm in diameter [77]. The
discharge power is less than ~ 3 W. The working temperature
of a plasma source is from 24 to 32 °C during plasma treat-
ment. The distance between the two electrodes of a C-DBD
can be changed from 0.1 mm to several centimeters. This can
also be used for the generation of facing DBD discharged
plasma by placing these two C-DBD panels opposite each
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other. Once gas is introduced through the peripheral guiding
hole with a high voltage applied, the discharge is fired on the
dielectric material, as shown in Fig. 2b.
The DBD-NBP can be applied for cancer cell death, skin
care, wound and burn treatment, agricultural and marine
products storage technology, semiconductor surface treat-
ment and environmental improvement for air, water and
soil. NBP devices operating at atmospheric pressure must
regulate ozone, which should be less than 0.05 ppm in living
environment [78, 79]; moreover, there must be no electric
shock and no heat generated during plasma treatment of liv-
ing tissue [5]. Especially a CDBD-NBP has made a remark-
able contribution to pathogen sterilizer in ambulances.
A CDBD-NBP approved as a very effective and efficient
sterilization and disinfection device for use in ambulances,
as well as many places, against COVID-19 throughout its
inactivation test.
2.2 Basic discharge physics in dielectric barrier
discharged (DBD) nonthermal biocompatible
plasma (NBP) operating at ambient
atmospheric pressure
Plasmas are rarely seen on the earth’s surface. We, therefore,
must make a plasma by using electrical breakdown. Figure 3
shows the schematic of discharge characteristics for coplanar
DBD (C-DBD) type of nonthermal biocompatible plasma
(NBP) with a gap distance d between the anode and cathode.
Seed electrons in a space with an electric field E collide with
neutrals, ionizing and generating new electrons. Here the
ionization rate α is called as the first Townsend coefficient
for ionization, which can be expressed in terms of electrical
field E and the gas pressure P [75, 76, 80] as
E BP
( ) ( )
𝛼 = AP exp − , (1)
P E
where A is a coefficient related to the ionization cross sec-
tion, B represents an unique property of the gas character-
istics such as the ionization potential energy, and P is the
Fig. 3  Schematic of the
discharge characteristics for a
coplanar DBD (C-DBD) type
of nonthermal biocompatible
plasma (NBP) with a gap dis-
tance d between the anode and
the cathode
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gas pressure between the anode and cathode separated by
gap distance d. For neon gas, A and B are given by A = 4
and B = 100 [81]. The α represents the fraction of newly
generated plasma electrons caused by ionizing electron
collisions with neutral molecules as an electron travels
by one millimeter along the electric field. The breakdown
sustaining condition of a gas between the two electrodes
with their gap distance d is given by γ[M − 1] = 1, where
M = e𝛼d is the electron multiplication factor estimated at
the anode and γ represents the secondary electron emission
coefficient, called the Townsend second coefficient, due to
low energy ions bombarding the cathode. The secondary
electron emission coefficient γ, defined by 𝛾 = Ie Ii , which
/
is the emitted number of secondary electrons (Ie) from the
cathode per ion colliding with the cathode (Ii). Normally,
secondary electrons are generated by quantum mechanical
Auger neutralization processes from the cathode surface
when a low-energy ion whose energy is less than 100 eV
approaches the cathode surface [82]. After some simple
mathematics, the breakdown-sustaining condition can be
rewritten as α(E/p)d = ln(1 + 1/γ). If the ionization constant
α(E/p) is substituted into the gas breakdown property, the
breakdown voltage defined by VB = Ed can be expressed as
[75, 76, 80, 81]
BPd
VB (Pd) = [ ],
APd (2)
ln
( )
ln 1+ 𝛾1
where the breakdown voltage VB is a function of the Paschen
parameter Pd, i.e., the product of the gas pressure P and the
gap distance d between electrodes. The breakdown voltage
for different discharge gases will be accordingly different
from each other since their respective gas constant B and
cathode property A are different to each other with differ-
ent secondary electron emission coefficient γ, as shown in
Eq. (2). Figure 4a shows the secondary electron emission
coefficient γ versus incident neon ion energy on a cathode
coated with a MgO layer with a crystalline orientation of
Plasma bioscience for medicine, agriculture and hygiene applications 821
Fig. 4  a Secondary electron emission coefficient γ for a MgO-coated
cathode with respective crystalline orientations of (111), (200), and
(220) versus incident neon ion energy in the range from 50 to 200 eV
[83]; b breakdown voltage, i.e., Paschen curve, versus Paschen
(111), (200), and (220). The neon ion energy is controlled
to be in the range from 50 to 200 eV. For the (111) orienta-
tion of the MgO layer, the γ is about 0.1 at a neon energy of
50 eV [83]. Figure 4b shows a Paschen curve, the so-called
breakdown voltage with a secondary electron emission coef-
ficient γ = 0.1 in the calculations for different gap distances
of 40, 80, 150, 200, and 250 um versus gas pressure P for
a Ne discharge gas with anode or cathode width of w = 180
um and a L = 540 um for x-space limit of geometry in right
hand side of Fig. 3 from its origin at a central point [81].
The minimum breakdown voltage VM could be obtained
from Eq. (2) by differentiating Vb (Pd) with respect to Pd and
setting the derivative to be zero, from which VM is given by
( ) ( ) ) (
B 1 2.72 1
(
VM = 2.72 ln 1 + at(Pd)min = ln 1 +
A A
𝛾 𝛾
where (Pd)min is the value satisfying = 0 for VM. The
dVb
d(Pd)
breakdown voltage VB increases when the value of Pd is too
small, which is less than 1 Torr-cm. Also, according to the
Eq. (3), each gas has its own minimum breakdown voltage
VM [75, 76, 80]. The discharge and its minimum voltage for
the shorter gap distances are lower than those for longer one
at gas pressure larger than 50 Torr.
parameter Pd in units of Torr-cm for different gap distances from
d = 40 µm to 250 µm calculated for the coplanar DBD geometry of
Fig. 3 [81]
3 Diagnostics of atmospheric pressure
nonthermal biocompatible plasma (NBP)
3.1 Optical emission spectroscopy (OES)
measurement
Optical emission spectroscopy (OES) can be performed to
identify reactive species by using the emission characteris-
tics of the nonthermal atmospheric pressure plasma. For this
kind of work, the spectrometer must be calibrated for wave-
length measurements by using an Hg–Ar lamp. The OES
spectra could be obtained by using an optical fiber whose
diameter is about 400 um placed in front of the nozzle of the
) plasma plume through which the emitted light is led to a slit
for collection into a grating of the spectrometer.
The optical emission spectra (OES) of a soft plasma jet
(3)
and a surface discharged micro-DBD are represented in
Fig. 5a and b, respectively. Emission signals from the nitric
oxide gamma band (NO-γ) at 236, 246, and 258 nm, and
283 nm whose energies are in the range of 3.27–5.25 eV
[84] can be seen in both Fig. 5a and b. These are caused
by the collisions of energetic electrons or metastable atoms
with nitrogen molecules in air. Also, emission in the range
of 306–309 nm, which are caused by hydroxyl radical (OH)
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Fig. 5  OES emission profiles from a the soft plasma jet and b the surface discharged micro-DBD plasma
species, which are produced by dissociating of water mol-
ecules, being present in the ambient environment [85].
The nitrogen second positive system (N2 SPS) is strongly
observed at 296, 315, 337, 356, and 380 nm, etc.; moreover,
the nitrogen first negative system (N2 FNS) emissions are
weakly observed at wavelengths in the range of 390–440 nm
[7, 86, 87]. These emissions, which originate from the
excited nitrogen species, are caused by nitrogen molecules
both in the feeding gas and the ambient environment. In
addition to these, emissions from atomic oxygen (O) are
seen at 777 nm and 844 nm, and the emission at 656 nm is
from the hydrogen atom (Hα).
3.2 Measurement of the OH and the NO radical
density using optical absorption spectroscopy
Reactive oxygen and nitrogen species (RONS) generated by
a non-thermal atmospheric pressure plasma play an impor-
tant role in many industrial fields. Radical generation from
plasma sources is an important phenomenon for the develop-
ment of plasma equipment because air plasmas can generate
abundant RONS. The optical diagnostic technology of the
plasma is very useful to control the amounts of chemical
species. The ROS has a very short lifetime and is used to
sterilize the biomaterials [5, 45]. The oxygen molecule is
known to be critical for aerobic biological or physiologi-
cal processes but it is converted to reactive oxygen species
(ROS) about 5% of the time during the energy production
processes [3]. These Reactive species are extremely toxic to
cells and causing damage to other molecules and cell struc-
tures. The nitric oxide (NO), a type of reactive nitrogen spe-
cies (RNS), performs an essential role in cell stimulus in
human body organs [88]. Many researchers have measured
these radicals by using optical absorption spectroscopy.
The hydroxyl radicals OH of ROS in a nonthermal
atmospheric pressure plasma can be diagnosed by using the
ultraviolet (UV) absorption spectroscopy. In Refs [86, 87
and 89], the experiment use of the ultraviolet absorption
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E. H. Choi et al.
spectroscopy is shown. This system consists of a UV lamp,
which is a Hg lamp whose power is 0.5 W centered at a
wavelength of 306 nm and a plano-convex lens which trans-
mits wavelength from ultraviolet to infrared [86]. The UV
light passes through and is focused by a plano-convex lens
with a diameter of 200 um in the plasma jet measurement,
the absorption that occur at 309.2 nm, for the OH species
[86]. The hydroxyl OH radical densities produced from air,
argon, nitrogen, or any kind of gas plasma can be investi-
gated by using Lambert–Beer’s law. The intensity for the
incident and the transmitted light passing through a plasma
space whose thickness is x are denoted an Io and Iv, respec-
tively, and the density of the hydroxyl radical OH species
produced by the plasma jet is given by [86, 87, 89, 90]
( )
1 I
N=− ln 𝜈 (4)
𝜎⋅x I0
where N is the density of hydroxyl OH radicals, and σ is the
molecular cross-sectional area, which is about 6 × ­10−7 ­cm2
for OH species [86]; x is 0.3 cm. The hydroxyl radical OH
density can be obtained from the experimental measurement
of Iv / Io, i.e., the ratio of the transmitted intensity to incident
intensity, by using Eq. (4).
Figure 6a shows plots of the UV absorption profile, rep-
resented by the black line, caused by the OH radical species
for a nonthermal Ar plasma jet whose gas flow rates range
from 80 to 300 sccm versus wavelength [86]. Also, the ref-
erence UV lamp (Io) and emission profiles from OH radi-
cal species in the nonthermal plasma without UV incidence
are indicated by red and blue lines, respectively, in this fig-
ure. The strong OH absorption profiles appear at 309.2 nm,
as shown in the dotted box, while no absorption is seen
at ~ 307 nm. In this experiment, the absorbed signals appear-
ing at ~ 317 nm have nothing to do with OH radicals. The
absorbed UV profiles (black line) in Fig. 6a can be obtained
by subtracting the emission profiles of OH radical (blue line)
in plasma from the transmitted UV intensity (Iv) passing
Plasma bioscience for medicine, agriculture and hygiene applications 823
Fig. 6  a UV absorption profile caused by the OH radical species
(black) versus the wavelength. Reference UV lamp profile (Io) versus
the wavelength without a nonthermal atmospheric pressure plasma
jet (red). Emission profiles from the plasma versus the wavelength
through the atmospheric pressure plasm jet [86]. The UV
emission and absorption profiles at 309.2 nm are denoted
by circles in the blue line and by the dotted box in the black
one, respectively, as shown in Fig. 1a [86]. The transmission
ratio (Iv/Io) for the UV lamp and produced by OH radicals,
can be used to estimate the OH density or concentration at
the absorbed wavelength of 309.2 nm. Figure 6b shows the
OH radical density at 2 mm above the interfacial surface
versus the Ar gas flow rate for rates from 80 to 240 sccm
under a low electrical power of 15 W with the driving fre-
quency of 22 kHz [86]. In this experiment, the OH density
reaches a maximum value of 2.6 × ­1015 ­cm−3 for a gas flow
rate of ~ 150 sccm, and it rapidly decrease to 6.0 × ­1014 ­cm−3
for a flow rate of ~ 250 sccm. Other groups also reported OH
densities of (0.3 ~ 7.5) × ­1015 ­cm−3 for low contents of water
molecules less than 3% for operating gases of He, ­N2, and
­N2/O2 mixtures at a microwave frequency 2.45 GHz and at
a RF frequency of 13.56 MHz with electrical power larger
than 100 W [85, 89, 90].
When a non-thermal atmospheric pressure plasma come
into contact with the water surface, the OH density increases
drastically to 6.55 × ­1016 ­cm− 3 at an appropriate applied volt-
age [86]. For the diagnostics of a nitrogen radical species such
as ­NOx, many researchers have generally used cavity-enhanced
absorption spectroscopy (CEAS) and Fourier transform infra-
red (FTIR) spectroscopy [91–93]. FTIR is a very useful radical
analyzer based on absorption spectroscopy, and a CEAS has
been employed in parallel with FTIR [91] for measurement
of radical species by including a multipath cavity cell with
a high reflection mirror [92] because FTIR spectroscopy has
low sensitivity. CEAS has a higher sensitive absorbance of
­10−7 ~ ­10−9, as compared with 1­ 0−3 ~ ­10−5 for FTIR. Also,
from hydroxyl OH radical species (blue) without UV incidence. b
Hydroxyl OH radical density on the region of the water surface in
contact with the plasma, that is 2 mm above the interfacial region,
versus the argon gas flow rate ranged from 80 to 240 sccm [86]
CEAS has a higher selectivity and a faster response time
[91–93] and is well adapted for the high‐speed dynamics of
RONS in a plasma gas. Hence, CEAS is suitable for in-situ
analysis and real‐time detection [93]. For nitrogen dioxide
­(NO2) measurements, the visible BBCEAS method can be
used; however, for nitric oxide (NO), CEAS should be used
with a mid‐infrared laser for diagnostics of infrared‐active
molecules, whose spectral range is between 3 and 20 μm [94].
The ­NO2 absorption can be measured by using ultraviolet and
visible light sources such as LED and Xe or Hg arc lamps. The
visible absorption band of ­NO2 includes the band of the elec-
tronic transition in the molecule [95]. However, the vibronic
absorption band of these NO species is located around 5.26 μm
and the absorption profile can be detected using a quantum
cascade laser (QCL). This QCL, which can be adjustable to
a specific wavelength of the laser, has been widely used for
NO absorption measurements. QCLs have the most suitable
characteristics for absorption spectroscopy. For these rea-
sons, ­NO2 and NO measurements can be carried out by using
the BBCEAS and the QCL–CEAS techniques, respectively.
In this review, we report the measurement of ­NO2 and NO
generated in a nonthermal air plasma jet and obtained using
BBCEAS with LED and CEAS with QCL, respectively. For
the measurements of the N ­ O2 and the NO densities, a visible
LED (light‐emitting diode) whose wavelength is 660 nm and a
mid‐infrared LD (laser diode) whose wavelength is 5.2386 μm
are used, respectively. Radical densities can be calculated from
the transmission ratio by using the Beer-Lambert law, which
is obtained from the amount of laser intensity absorbed where
passing through the gas from the plasma jet in the optical cav-
ity of CEAS [96]. QCL–CEAS has generally been used to
measure the absorption of NO radical species. Especially, the
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824 E. H. Choi et al.

Fig. 7  a Transmission ratio of beam versus wavelength of QCL, and b the N
­ O2 and NO radical density versus duty ratio (%) [96]

­ O2 density has a value of ~ 2.5 × ­1016 ­cm−3 in our air plasma

N N2 (B) + N2 (X) → N2 (A) + N2 (X). (9)
jet. The NO density has a value of ~ 4 × ­1015 ­cm−3 [96]. The
transmission ratio was measured in the range of the QCL’s Also, ­N2 (A) under goes wall deactivation [98],
tunable wavelength to find the maximum absorption wave-
length of NO. Figure 7a shows the absorption wavelength of N2 (A) →wall N2 (X) (10)
5.2386 μm at the graph’s first peak [96]. Figure 7b was the
and the two excited states, A 3Ʃu+ and B 3Πg, have the effect
graph for the relation of the N
­ O2 to the NO radical densities
of spontaneous emission [98]:
according to the pulse duty ratio [96].
wall
N2 (B) → N2 (A) + hf , (11)
3.3 Measurement of the electron temperature
and density
N2 (C) → N2 (B) + hf , (12)
We refer to the nitrogen collisional radiative model of Xi- where f is the emission frequency. The processes of Eqs. (11)
Ming Zhu and Yi-Kang Pu [97, 98] to obtain the electron and (12) cause the spectra of ­N2 SPS (second positive sys-
temperature and density in an air plasma jet. The processes tem) and ­N2 FPS (first positive system) spectra, respectively.
involve the ground state X 1Ʃg+and the excited states A 3Ʃu+, Additionally, when the pressure is above 30 mTorr, we
B 3Πg and C 3Πu [97, 98]. The main processes are electron have to consider the energy pooling reaction and collision
impact excitation [97, 98], quenching by nitrogen atoms [98]. For the determination
e + N2 (X) → e + N2 (A), (5) of an approximate electron temperature in an atmospheric
pressure plasma, we only included energy pooling reaction
as follows [97, 99]:
e + N2 (X) → e + N2 (B), (6)
N2 (A) + N2 (A) → N2 (B) + N2 (X) (13)
e + N2 (X) → e + N2 (C), (7)
N2 (A) + N2 (A) → N2 (C) + N2 (X) (14)
­ 2 (X) is a nitrogen molecule in the
where e is the electron, N
ground state, and ­N2 (A), ­N2 (B), and ­N2 (C) are excited Therefore, we can build up the new balance equation
nitrogen molecules for A, B and C states, respectively [97, for the three excited states included in the energy pooling
98]. A nitrogen molecule in the A state can collide with are reaction in the original balance equation of Ref. [98]. In
in the ground state: the case of ­N2 (A),
N2 (A) + N2 (X, 5 ≤ v ≤ 14) → N2 (B) + N2 (X), (8) ne ng QA + AB nB + kBX ng nB = kwall nA + kAX nv nA + 2n2A (kAAB + kAAC )
(15)
where v is the vibrational quantum numbers [98]. The
excited ­N2 (B) can be quenched by collisions with nitrogen In the case of ­N2 (B),
molecules in the ground state:

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Plasma bioscience for medicine, agriculture and hygiene applications 825

ne ng QB + kAX nv nA + Ac nc + n2A kAAB = kBX ng nB + AB nB (16)

In the case of ­N2 (C),

ne ng QC + n2A kAAC = AC nC (17)

The ne, nA, nB, and nC are the densities of electrons and
excited states A 3Ʃu+, B 3Πg, and C 3Πu, respectively. The
nv is the neutral gas density for the vibrational tempera-
ture and the ng is the neutral gas density, which depends
on the gas temperature. The kAX and kBX are the rate coef-
ficients for the collision processes in Eqs. (8) and (9) [98].
The kAAB and kAAC​ are the rate coefficients for the process
of exiting to upper states B 3Πg and C 3Πu after the colli-
sion between excited states A 3Ʃu+ in Eqs. (13) and (14)
Fig. 8  Electron temperature and density for the line ratio curve
[97–99]. Also, in Eq. (15), the kwall is the rate coefficient of between ­N2 SPS and FPS
wall deactivation, which is achieved by the diffusion model
with the coefficient of wall reflection [98]. The QA, QB, and
QC are the rate coefficients for electron impact excitation
from the ground state to excited states A 3Ʃu+, B 3Πg, and C
3
Πu, respectively [98, 100]. The AB and AC are the transition
probabilities of excited states B 3Πg and C 3Πu [98]. There-
fore, we can obtain three 2­ nd-order simultaneous equations
in three unknowns the excited molecule densities nA, nB, and
nC. Equations (15)–(17) can be arranged as follows:
2n2A (kAAB + kAAC ) + (kwall + kAX nv )nA + (AB + kBX ng )nB − ne ng QA = 0
(18)
n2A kAAB + kAX nv nA −(AB + kBX ng )nB + Ac nc + ne ng QB = 0
(19)
n2A kAAC − AC nC + ne ng QC = 0. (20)

These excited nitrogen molecule densities can be calcu-

lated simply by using a software library called the solve
function in MATLAB or Python. In addition, we can express
the density of each excitation molecule in terms of these
electron temperatures and densities by assigning an arbitrary
electron temperature and density to Eqs. (18)–(20). The ratio
of molecule densities for nitrogen SPS and FPS can be writ-
ten as follows [98]:
) A n
R kTe , ne = C c
(
AB nB
The values of Eq. (21) have been compared with the
measured emission line ratio between ­N2 SPS and FPS. ­N2
SPS emission lines have been selected at wavelengths of
295.32, 313.60, 315.93, 337.13, 353.67, 357.69, 371.05,
375.54, 380.49, 389.46, 399.84, and 405.94 nm [101]. Also,
­N2 FPS intensity was for a wavelength of 654.49 nm [101].
The measured value R = IC IB in Fig. 8 is the line ratio
/ 3.4 Plasms gas temperature measurement
between the one value of twelve lines for ­N2 SPS and the
line for FPS. Twelve curves of line ratios can be obtained
Fig. 9  Curve for finding the crossing point of two curves for
the electron temperature and density of I295.32 nm/I654.49 nm and
I353.67 nm/I654.49 nm. [101]
for each arbitrary electron density. The electron temperature
for a specific electron density can be obtained by finding
the value at the curve of Eq. (21) that matches measured
value of the emission line ratio. Therefore, we can express
(21) the curve of electron temperatures for arbitrary electron
densities. Figure 9 shows the cross point for two curves in
the case of I295.32 nm/I654.49 nm and I353.67 nm/I654.49 nm. The
determined electron temperature and density are 0.95 eV and
2.5 × ­1015 ­cm−3, respectively, in the Fig. 9. We can calculate
the electron temperature and density in a air plasma by using
this method.
The plasma gas temperature is fundamentally important
for understanding the characteristics of the nonthermal

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826 E. H. Choi et al.

atmospheric pressure (N-DBD) plasma used in most clini- The intensity IJ'J" of a particular rotational transition
cal medicines. The Boltzmann distribution of rotational N ­ 2 J" → J″ in this SPS is given by [30]
molecular levels can be used to measure their gas temper- ( ( ))
ature in N-DBD plasmas [102–104] by using the optical Er v� , J �
(22)
( �� )
Ij� j�� = Av� v�� SP J nc exp −
emission spectroscopy [102–104]. Most rotational and vibra- kTrot
tional gas temperatures can be obtained by using the second
positive system (SPS: ­N2 ­C3IIu → ­N2 ­B3IIg) [102–104] of ­N2 where Avv′ is transition amplitude, Sp(J) = 6 J″–10/J″ is the
molecular levels rather than the first positive system (FPS: Hoenl-London factor corresponding to the rotational angular
­N2+ ­B2Σu+ → ­N2+ ­X2Σg+). Figure 10 shows the transitional momentum quantum number J″, and E ­ r (υ′, J′) is the rota-
energy diagram for ­N2 SPS (­ C3IIu → ­N2 ­B3IIg) and ­N2 FPS tional energy for vibrational number υ’ and rotational num-
­(N2+ ­B2Σu+ → ­N2+ ­X2Σg+) (a) and the optical emission spec- ber J’ given by
tra of the N-DBD plasma jet, where ­N2 SPS (297–400 nm) )2
Er v� , J � = Bv J � J � + 1 − Dv J � J � + 1 (23)
( ) ( ) (
and N 2 FPS (550–800 nm), as well as the NO-γ band
(213–258 nm) are observed [105]. Vibrational spectra of
the ­N2 SPS and their magnified rotational spectra belonging where ­Bυ and ­Dυ are rotational term for vibrational number
to the 0–0 vibrational band are shown in (c) around 337 nm υ, ­J’ and J­ ’’ are the rotational quantum numbers for the tran-
for ­N2 ­C3IIu (v = 0) → ­N2 ­B3IIg (v = 0), as indicated by the sitions (υ = 0, J′)→( υ = 0, ­J″), k is the Boltzmann constant,
dotted box.

Fig. 10  a Transitional energy diagram N2 SPS ­(C3IIu → ­N2 ­B3IIg) and nified rotational spectrum in the 0–0 vibrational band around 337 nm
­N2 FPS ­(N2+ ­B2Σu + → ­N2+ ­X2Σg+). b Optical emission spectra of the of ­C3IIu (v = 0) → ­B3IIg (v = 0), as in the dotted box (c) [105]
­N2 SPS and the N2 FPS. c Vibrational spectra of the N­ 2 SPS and mag-

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Plasma bioscience for medicine, agriculture and hygiene applications 827

and Trot is the rotational temperature. For an estimate of the for a nonthermal air plasma plume above 1 mm above the
rotational temperature, Eq. (6) can be rearranged as: surface of the water (Fig. 11).
�
The vibrational gas temperature can also be estimated by
B hc ( selecting quantum levels of the SPS corresponding to the ν–ν′
( )
I � ��
In − J J �� = − 𝜈 J� J� + 1 ) + C (24)
)
Sp (J ) kTrot vibrational band of N­ 2 ­(C3 Πu) (v)—B3Πg (v′). The intensity
of a particular vibrational band in this SPS is given by [106],
where h is Plank’s constant, c is the speed of light, and C is
constant. Figure 12 shows a plot of Eq. (24) versus rotational
( )
I𝜈𝜈 � E − E0
ln =C− 𝜈 (25)
energy given by Eq. (23) with rotational quantum number 𝜈𝜈𝜈 � A𝜈𝜈 � kTvib
J’. The rotational temperature Trot of the plasma gas mol-
ecules can be determined from the reciprocal value of the where I𝜈𝜈 ′ is emission intensity of transition 𝜈 → 𝜈 ′ , measured
slope, Bυ’hc/(kTrot), of the resulting linear plot of the left- from experimentally, 𝜈𝜈𝜈 ′ is the transitional frequency, A𝜈𝜈 ′ is
hand side of Eq. (24) versus the rotational energy and Er. transitional amplitude, and E𝜈 is the vibrational energy for
From this, throughout repeated experiments, the rotational quantum number 𝜈 given by
temperature Trot of the plasma gas is estimated to be 807 K

Fig. 11  a Rotational temperature Trot of the plasma gas molecules can
be determined from the reciprocal slope of a linear plot of the left-
hand side of Eq. (8) versus the rotational energy and Er. This plasma
gas temperature is estimated to be 807 K for a nonthermal air plasma
plume 1 mm above the water’s surface; b Estimate of rotational gas
temperature at a position of 12 mm below the nozzle of the quartz
tube by using a Boltzmann plot of the nitrogen second positive sys-
tem (SPS) (0–2: vibrational band; peak wavelength: 380.4 nm) (blue
liner), which yield 325 K at an Ar flow rate of 250 sccm. Also its
comparison with the temperature taken from a fiber optic thermom-
eter (red one) [104]

Fig. 12  a A schematic showing how ions, electrons and neutral particles, as well as UV radiation, contribute to the production of reactive oxy-
gen and nitrogen (RONS) species in gas and water phases during the NBP discharge [21], and b their approximate half-lives [21]

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828
(
1
)[
E𝜈 (eV) = 1.24 × 10−4 + 𝜈 𝜔e (cm)−1 . (26)
]
2 emits UV light whose wavelength is in the range from 200 to
The vibrational temperature of Tvib of the plasma mol-
ecules can be determined from the reciprocal value of the
slope of the resulting linear plot of the left-hand side of
Eq. (25) versus energy difference E𝜈 − Eo. From this, the
vibrational temperature kTvib of the plasma gas is measured
to be 0.2–0.8 eV for nonthermal air plasma plume above
1 mm above the water’s surface [104]. Because the density
of excited nitrogen molecules in the excited energy bands C,
B, and A are up to ~ ­1017/cm3, the ratio of excited molecules
with both rotational and vibrational temperatures to those of
ambient air molecules is estimated to be about 1% where the
remaining 99% of most ambient molecules are at room tem-
perature and quickly absorb energy due to the rotational and
the vibrational temperatures of these 1% excited molecules,
resulting a cool of the temperature, as can be felt when your
hand touches this DBD plasma. Hence, we call this kind of
plasma as a nonthermal, cold, or cool plasma even though
the rotational and the vibrational temperatures are somewhat
higher than room temperature.
4 Plasma interaction with water
Most plasma induced reactive oxygen and nitrogen spe-
cies (RONS) are generated when the NBP interacts with
the water’s surface or water molecules in the ambient air or
within tissue. Here, much physics and chemistry are involved
and they are important for understanding the mechanism
underlying plasma treatment of wounds or cancer because
water is a major component of plasma treatment and it is the
simplest hydrated environment in plasma-liquid interactions.
Within the simplest water-based model, the mechanisms of
why and how NBP generates RONS in water or solution is
somewhat solved, but still remain unclear. This is due to
the technical difficulty in in-situ measurements of highly
reactive molecules generated by the NBP in water, such
as the hydroxyl radicals (OH•), superoxide anions ­(O2*−),
nitric oxide (NO), peroxynitrite (­ ONOO−), and other radi-
cal species.
Figure 12a represents the schematics showing how ions,
electrons and neutral particles as well as UV radiation,
generate RONS in gas and liquid phases during a plasma
discharge and Fig. 12b their approximate half-lives [21].
Energetic electrons formed in the discharge region can col-
lide with neutral atoms like nitrogen, oxygen, water vapor,
etc. and dissociate them. The primary species, OH, NO, O,
­O2*−, 1O2, and N­ 2+, are generated in the discharge region.
These primary reactive species [5, 53, 54, 60] in the gas–liq-
uid interface with their half-life times ranging from nano or
microseconds (O and OH), milliseconds ­(ONOO−, ­O2*−),
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E. H. Choi et al.
seconds (NO, 1O2), to several minutes ­(H2O2, ­NO2−, ­NO3−),
400 nm inducing the secondary reactive species in the water.
We call this kind of the simultaneous secondary RONS pro-
duction mechanism in water, it as plasma ultraviolet (UV)
photolysis [7, 84–86].
4.1 Plasma ultraviolet photolysis and molecular
transport of plasma reactive species
The RONS chemistry induced by the plasma UV radiation
originating from NBP is not homogeneous inside the water,
where highly reactive molecules are depleted in regions far
from the from the water-plasma interface. The physical and
chemical mechanisms for generating plasma-induced reac-
tive species in water should be synergistic combination from
simultaneous RONS production of plasma UV photolysis
and subsequent gas-region RONS diffusion into the water
[55, 96–98]. Some significant advances in the understanding
of the mechanisms have been achieved and are summarized
[107, 108].
The generation OH and H ­ 2O2 in a plasma UV photolysis
with an electrical discharge power of 4.9 W and a driving
frequency of 35 kHz, can be visualized inside DI water by
irradiating of UV light onto the water surface. The diag-
nostic methods for these species will be explained. Gener-
ally, UV light can be produced by using mercury lamp or
any other sources whose wavelength is centered at 306 nm.
Terephthalic acid (TA) has been put into the water for visual
check of OH production in the water or biosolutions during
UV bombardment [109]. Figure 13a confirms that OH spe-
cies can be generated inside the DI water and PBS, when the
liquid’s surface with/without quartz filter is placed at 1 mm
below the water’s surfaces is irradiated with UV light with
its energy entered at 4 eV. Because blue color is observed.
This UV light can pass the filter and propagate into the water
for excitation and dissociation of it, resulting in OH genera-
tion. A visual observation of OH species inside the water
can be done either by using a red color filter (right side in
Fig. 13a) or an irradiation of incandescent light onto the
water. No color change for these cases.
Figure 13b shows the generation of H ­ 2O2 inside the DI
water either by UV light or Ar plasma jet bombardment at
an electrical power of 4.9 W and a driving frequency of
35 kHz. Here, 0.01 M ammonium metavanadate ­(NH4VO3)
is put into the DI water and PBS to observe ­H2O2 production
visually when the color changes to orange. Furthermore,
the ­H2O2 generation inside the DI water can be confirmed
by using a quartz filter when Ar plasma is bombarding the
surface. The quartz filter, which is placed just on the water’s
surface, can screen plasma electrons, ions, and neutral parti-
cles so that only the plasma UV emission, which are caused
Plasma bioscience for medicine, agriculture and hygiene applications 829
Fig. 13  a Visual observation of OH generation inside DI water and
PBS with/without TA when a UV mercury lamp is used for the irra-
diating; b visual observation of H
­ 2O2 generation inside the DI water
either by UV irradiation or plasma bombardment of the water with/
from the excited ROS species, can pass through the filter to
propagate into the water.
The ­H2O2 and OH species can be generated simultane-
ously inside DI water and PBS by using plasma UV irra-
diation. Here the OH density inside the water, produced by
plasma UV photolysis, has been observed to have a correla-
tion with the H­ 2O2 concentrations. Figure 13c shows there is
a strong correlation between the OH and the H ­ 2O2 concen-
trations in the DI solution, where the OH density is increased
from 1.3 × ­1016 ­cm−3 to 2.3 × ­1016 ­cm−3 as the H ­ 2O2 con-
centration is changed from 0 to 0.8%, respectively. Also,
the OH densities (DI, PBS) reach maximum values of (4.2,
1.9) × ­1016 ­cm−3 and (0.8, 0.1) × ­1016 ­cm−3 at depth of 2 mm
and 6 mm, respectively, below the water’s surface, as shown
in Fig. 13d at a gas flow rate of 250 sccm [110].
4.2 Plasma parameter characteristics
at plasma‑water interfacial region
In this section, plasma parameter variations for the interaction
between a plasma jet and deionized surface are introduced.
Atmospheric pressure Ar, air, N ­ 2 plasma jet and deionized
water can be used for this. A cuvette filled with DI water is
placed 10 mm below the quartz tube nozzle [111]. Optical
without quartz filter located just on the DI water; c OH density versus
the external ­H2O2 concentrations in DI water at a depth of 4 mm for
an Ar plasma jet; d density of OH versus the depths in the DI and the
PBS solutions produced by an Ar plasma jet [110]
emission spectroscopy is performed with a spectrometer with
an optical fiber. The optical fiber is installed behind of quartz
plate to avoid arcing. A deuterium lamp and a high-resolution
spectrometer are installed with lens for absorption spectros-
copy of the hydroxyl radical. The optical diagnostics is focused
on a plasma bullet located 5 mm below the nozzle.
In this experiment, the length of the plasma jet was
observed at least 10 mm from the nozzle regardless of the pres-
ence or absence of water. The measured plasma discharge volt-
age (black line) and current (blue line) are shown in Fig. 14a
as functions of time. The rms value of the driving voltage
was measured as 2.3 kV, and the plasma discharge current
was measured as 10 mA. The plasma bullet current (red line)
is measured by Rogowski coil as shown in Fig. 14a and it
is found to be 70 mA. The average power dissipation by the
plasma discharge (PDis) can be calculated using the voltage
(v(t)) and the current (i(t)) from the grounded copper tape by
using [112]
T
T∫
1
PDis = v(t)i(t)dt, (27)
0
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830
Fig. 14  a Plots of the voltage (black line) and the current from the
pickup coil (blue line) and Rogowski coil (red line) versus time at
2.3 kV with DI water, b dissipated power of Ar APPJ with (red-circle
line) and without (black-square line) DI water versus applied volt-
where T is the period of voltage. The dissipated power of
both with and without DI water are shown in Fig. 14b. They
are not very different from each other. However, a signifi-
cant difference between them appears to occur at voltage
above 2.3 kV. Above 2.3 kV, the plasma bullet’s current on
DI water starts to increase because the DI water’s surface
serves as a ground. Therefore, the dissipated power in the
region of the grounded copper tape region of with DI water
is saturated. The velocity and the peak current values of
the plasma bullet as functions of the applied voltage are
shown in Fig. 14c. A Rogowski coil is used for plasma bul-
let velocity measurements, and its speed was calculated by
time difference (Δt) and distance (ΔL) between the current
in the pickup coil and that in the Rogowski coil. The plasma
bullet currents with and without DI water are found to be
in the ranged from 7.6 to 57.4 mA and 3.6–10.1 mA as a
function of applied voltage, respectively, and the plasma
bullet’s speeds are found to be ranged from 8.5 to 9.9 km/s
and 12.7–13.9 km/s as a function of applied voltage, respec-
tively. When a voltage of 2.3 kV is applied, the plasma bul-
let’s current increases as shown in Fig. 14c due to the strong
electric field between the plasma bullet and the DI water
surface. Thus, the plasma current flows are divided into two
parts: the current in the grounded copper tape and the cur-
rent in the plasma bullet directed to the water surface. The
bullet’s current is increased with DI water because the DI
water’s surface serves as a ground that makes the electric
field between them stronger. The water vapor for the DI
water can reduce the plasma bullet’s velocity due to colli-
sions between plasma electrons and water molecules. The
chemical reaction for the generation of OH radicals can be
explained as the dissociation of water molecules. In the DI
water’s surface, vaporized water molecules in the gaseous
state can increase OH production through collisions with
metastable Ar and O atoms and can increase electrons in
plasma bullets for interaction [113–115]. The emission
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E. H. Choi et al.
age, c speed of the plasma bullet (red-star line and orange-circle line)
and the current from the Rogowski coil (blue-square line and purple-
triangle line) with and without DI water versus the applied voltage
[111]
intensities of Ar and O as functions of applied voltage pro-
vide the evidence of this.
Figure 15 shows optical emissions from a plasma bullet
5 mm below the nozzle. The OH band (306–312 nm), ­N2
second positive system (337 nm, 380 nm), O (777 nm) and
Ar (696–965 nm) are measured both with or without DI
water and the results are shown in Fig. 15a. The intensities
of Ar emission, two emission lines at 912 nm and 696 mn,
are observed as a function of applied voltage as shown
in Fig. 15b. There emission intensities with or without
DI water were increased steadily until 2.3 kV. Especially,
the maximum value was reached with DI water, which is
increased significantly from 2.3 kV. Water vapor due to the
interaction between the plasma bullet and the DI water’s
surface increases collisions with metastable Ar atoms,
resulting in a loss of Ar metastable energy. Therefore,
when a voltage of 2.3 kV or more is applied, the intensity
of Ar emission from DI water decrease. The O atom can
be obtained from the dissociation of the O ­ 2 molecule by
metastable Ar atoms and electrons [91, 93].
Ar∗ + O2 → Ar + O + O (28)
e− + O2 → e− + O + O(1 D) (29)
The O I emission intensities are measured as func-
tions of the applied voltage and the results are shown in
Fig. 15c. These emissions increase with increasing applied
voltage with or without DI water, especially with DI water
at voltage beyond 2.3 kV. According to the results for the
plasma bullet’s peak current in Fig. 14c and the Ar emis-
sion intensity in Fig. 15b, O atoms appear to arise during
the processes described in Eqs. (28) and (29) by the elec-
trons of the plasma bullet and metastable Ar atoms.
Plasma bioscience for medicine, agriculture and hygiene applications 831

Fig. 15  a Optical emission spectroscopy results for a APPJ with triangle line) with and without DI water as a function of the applied
(red line) and without (black line) DI water at an applied voltage of voltage. c O I (777 nm) emission intensities with (red-circle line) and
2.3 kV. b Metastable Ar emission intensities at 912 nm (red-star line without (black-square line) DI water versus the applied voltage [111]
and orange-circle line) and 696 nm (blue-square line and purple-

Fig. 16  a Electron temperature versus applied voltage, and b electron density versus applied voltage [111]

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832
Electron temperature in the plasma bullet has been meas-
ured by Ar collisional radiative model and the results are
shown in Fig. 16a, and is found to be 1.36–1.06 eV with DI
water and 1.38–1.42 eV without DI water. This shows that a
decrease in the electron temperature with DI water is caused
by a water vapor increase. At a 2.3 kV applied voltage, an
increase in the water vapor on the DI water’s surface leads
to a decrease in the mean free path for electron collision,
which leads to electron temperature decrease. At the same
time, measured electron densities with DI water are found
to be 1.02 × ­1015 ­cm−3 to 8.62 × ­1015 ­cm−3 with DI water
and 6.58 × ­1014 ­cm−3 to 1.33 × ­1015 ­cm−3 without DI water
as shown in Fig. 16b. The increase in the electron densities
with DI water can be explained by the presence of water
vapor and the following processes [116]:
e− + H2 O → H2 O+ + 2e−
e− + H2 O → OH+ + H + 2e−
In addition, it can be explained by an increase in the oxy-
gen emission intensity, are found in Fig. 15c, due to Eq. (29)
caused by an electron density increase.
Figure 17a shows that the absorption profile for OH
lines in the range from 306 to 312 nm at a 2.3 kV applied
voltage with DI water. The corresponding OH density
results are plotted against the applied voltage in Fig. 17b.
In this experiment, 309 nm is selected in the absorption
spectrum to measure the density of OH radicals. The
results obtained with or without DI water increase with
the increasing applied voltage. Especially in the presence
Fig. 17  a Absorption profile at 308 nm, and b OH densities with/without DI water versus applied voltage [111]
13 Vol.:(0123456789)
E. H. Choi et al.
of DI water, at an applied voltage of 2.3 kV, it rapidly
increased to 6.55 × ­1016 ­cm−3, which is about seven times
higher than the value at 2.2 kV. This increase in the OH
radical with DI water can be explained by dissociation of
water molecules by Ar atoms, O atoms and electrons [85,
91–93] as follows:
Ar∗ + H2 O → OH + H + Ar (32)
O(1 D) + H2 O → 2OH (33)
e− + H2 O → OH + H + e− (34)
The reaction rate coefficients of Eqs.
(32)–(34) are 7.8 × ­1 0 –10 ­c m 3/s, 2.0 × ­1 0 –10 ­c m 3/s, and
(30) (1.68Te − 1.23Te2 + 2.19Te3 ) × 10−10 ­cm3/s, respectively, at
a gas temperature ­Tg ≈ 300 K [114–116].
(31) Because the emission intensity of metastable Ar atoms
in DI water at applied voltage above 2.3 kV is higher
than those obtained up to 2.3 kV, the reaction shown in
Eq. (31) is one of the main reasons for the increase in the
OH densities shown in Fig. 17b. The reaction depicted in
Eq. (33) for the generation of OH radicals can be enhanced
by increasing the number of O atoms as shown in Fig. 15c.
The rate coefficient of Eq. (34) with DI water is reduced
from 5.41 × ­10–10 ­cm3/s to 4.92 × ­10–10 ­cm3/s by decreasing
the electron temperature from 1.35 to 1.30 eV, whereas
the electron density increases from 4.12 × ­1 0 15 ­c m –3 to
6.47 × ­1 0 15 ­c m –3 at applied voltages ranging from 2.2
to 2.3 kV. As a result, the OH radical species can be
increased by the reaction depicted by Eq. (34) when a volt-
age of 2.3 kV is applied.
Plasma bioscience for medicine, agriculture and hygiene applications 833
5 International standard for plasma medical
equipment
5.1 Measurement of the plasma current
The current between the plasma and the skin is also one of
the biological effects of a plasma. Among the conventional
medical devices are devices that are expected to have a ther-
apeutic effect caused by an electric current on the patient.
Safety limits and measurement methods for the current
through the patient are specified in IEC 60601-1-1. The safe
value for the patient current given in IEC 60601-1-1 is 100
uA [117]. Sensitive people may feel uncomfortable when
sensing the current even when the tolerance is satisfied. The
detection threshold is individual and depends on age and
gender. From a physiological point of view, the tolerances
given in IEC60601-1 do not cause health problems [118].
The leakage current for a soft jet can also measured by
among a copper plate target (40 × 40 × 5 ­mm3) connected to
a test device (UNIMET® 800ST, BENDER), as shown in
Fig. 18a, according to the axial distance [119]. As shown in
Fig. 18b, the current measured up to only 2 mm from the soft
jet nozzle, and no more current is measured beyond 3 mm
from the nozzle because the lower measurement limit of the
instrument is 1 uA.
5.2 Measurement of ozone and NOx
Atmospheric pressure plasma discharge generates reac-
tive oxygen species (ROS) such as ozone and OH, H ­ 2O 2,
and reactive nitrogen species (RNS) such as NO and ­NO2
[119]. Reactive nitrogen oxygen species (RONS) gen-
erated by a plasma play an important role in biological
interaction [120, 121]. Some RONS have toxic properties
and require caution in biological applications. Control of
RONS produced by using an atmospheric pressure plasma
Fig. 18  Medical requirement
limitation measurement: a
plasma current and b plasma
current versus distance under
several off-time durations in a
discharge for a soft plasma jet
is essential. Safety standards and accurate measurement
methods for active species order to ensure the safety of
medical personnel and patients when atmospheric plasma
medical devices.
In general, ozone and ­NO2 are known as air pollutants
that have harmful effects on the respiratory systems of
humans and animals [122, 123] In the case of ozone, long-
term exposure has been reported to be associated with the
occurrence of asthma [124]. If the ozone concentration in
the atmosphere is more than 0.02 ppm, we can sense the
smell. The safety standard recommended by the Ministry
of Environment of Korea and ACGIH (American Confer-
ence of Governmental Industrial Hygienists) of the United
States is 0.05 ppm. It is said that no danger exacts below
this standard. If more than 0.44 ppm of ­NO2 is generated,
people can smell it. N
­ O2 gas may cause cardiovascular dis-
ease when exposed to it for a long time [117]. The safety
standard recommended by the Ministry of Environment of
Korea and by the ACGIH of the United States is 3 ppm,
which should not be exceeded for the health of humans
and animals. On the other hand, NO gas is known as an
essential substance for maintaining the homeostasis of the
human body [125, 126]. In particular, NO is an antibacte-
rial substance that plays an especially important role in
the immune system that protects the human body from
microorganisms [127].
The measurement method for RONS suggested in the
plasma medical device standard is to be from plasma gen-
erators are spread. Measured at three locations as shown
in the Fig. 19. Figure 19a presents a method for measuring
RONS from the front of the plasma generator [126].
Ozone can be measured in accordance with the measure-
ment distance from the nozzle of the soft plasma jet by using
a commercial device (200 series, aeroqual), as shown in
Fig. 20a. In Fig. 20b, ozone production from the soft plasma
jet is seen to be increasing with distance, and it is slightly
increased in accordance with the increasing of its off time.
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5.3 Measurement of the plasma plume temperature
at the target
When treating wounds, no damage should be caused by
heat from on atmospheric pressure plasma. Therefore, the
thermal energy in an atmospheric pressure plasma must be
measured. According to IEC 60601-1-1, the temperature of
medical devices should not exceed 40 ºC. However, a slight
increase in temperature can induce proliferation of living
keratinocytes [127]. Therefore, treatment with an atmos-
pheric pressure plasma can actively induce wound healing
and tissue regeneration if the temperature of plasma does
not exceed 40 ºC. At temperatures, protein denaturation and
membrane destruction are well known to occur.
The temperature of the plasma device is shown in
Fig. 21a. The results of measuring the temperature according
to the distance in the axial direction are shown in Fig. 21b.
The temperature profile of soft jet is shown in Fig. 21b

Fig. 19  Method for measurement the gas emission at different angles a 180º, b 90º, and c 45º

Fig. 20  Medical requirement

limitation measurement: a
ozone measurement and b
ozone concentration versus
distance under several off-time
durations in the discharge of a
soft plasma jet

Fig. 21  Medical requirement

limitation measurement: a axial
temperature measurement, and
b temperature versus distance
under several off-time durations
in the discharge of a soft plasma
jet

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Plasma bioscience for medicine, agriculture and hygiene applications 835
[128]. As shown in the figure, the temperature is 35–40 ºC at
7–10 mm from the jet tip. This distance becomes the recom-
mended distance for a soft jet and should be used on patients
at this distance to keep the plasma temperature below 40 ºC
[128, 129]. These temperatures are biologically safe and are
permitted by IEC60601-1-1.
6 Utilization of plasma bioscience for living
health
The healing mechanism of lesion tissues related to bacteria
infected and virus-infected living tissues under plasma treat-
ment can be clarified through basic and clinical research
by using food-borne and air-borne pathogens. Also, imple-
menting plasma medical devices for these kinds of medical
purposes is important. This requires an absolute coopera-
tion system between the two organizations for international
standards of food safety and plasma medical devices, joint
use of advanced research facilities, joint acquisition of intel-
lectual property rights that will lead the world, and synergy
of securing advanced technologies. In particular, the plasma
bioscience field shows promising features that could be
widely used in the prevention and the treatment of COVID-
19 [44, 130], which is pandemic around the world.
6.1 Plasma agriculture and food treatment
in fisheries
6.1.1 Plasma agriculture and fishery
Plasmas have an effect on seed germination. From ancient
times, there has been a story that grain grows well and har-
vests are good in summer when there is a lot of thunder and
lightning. All of them make sense from the perspective of
plasma bioscience. As shown in Fig. 22a, water treated by
Fig. 22  a Effect of treatment using NBP on the germination and growth of tomato seeds [26]. b Schematic view of DBD-NBP plasma treatment
on the reduction of S. aureus and B. cereus on black mouth angler fish [131]
a “thunderbolt” on tomato seeds, especially seeds sprinkled
with plasma-treated water for 15–30 min, grows better and
shows strong pathogen resistance [23–34]. This is due to an
increase in the growth hormone and the pathogenic hormone
resistance of plants when seeds are exposed to a plasma [26].
In addition, Staphylococcus aureus (S. aureus) has strong
resistance and is widely present in air and soil.
It lives on the skin of humans and animals, is easily con-
taminated through food, and can cause food poisoning and
sepsis. As shown in Fig. 25b, The NBP-DBD surface plasma
can control Staphylococcus aureus (S. aureus) and Bacillus
cereus (B. cereus) bacteria in dried anglerfish, which are
related to food poisoning [131]. More than 90% of S. aureus
and B. cereus killed when treated for about 30 min; the effect
is about 99% after about 1 h [131]. The important point here
is that the color coordinates do not change significantly, and
even the shape, taste, aroma, and overall water solubility of
dried anglerfish are not significantly affected. Therefore, in
the case of dried fish, if a large-area DBD-NBP plasma is
irradiated for about 30 min, a sterilization rate of 90% or
more can be expected, and no side effects are expected to
occur. Therefore, plasma treatment of dried seafood can be
considered during production, processing, and distribution
under conditions that sufficiently ensure food safety. Like-
wise, plasmas can be actively used in the production, storage
and distribution of agricultural and livestock products.
6.2 Inactivation of antibiotic‑resistant
superbacteria
In everyday life, people visit hospitals to receive medi-
cal treatment when they are sick or to hospitalized family
members. They can be infected indoors for no reason while
waiting for treatment, or during visits to the hospital. Also,
they can even be infected with antibiotic-resistant super-
bacteria in the hospital, putting them at risk. Infection with
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antibiotic-resistant superbacteria, which has a resistance to
methicillin as well as penicillin, is becoming a serious prob-
lem. Antibiotic-resistant bacteria include penicillin-resist-
ant Staphylococcus aureus (PRSA), gentamicin-resistant
Staphylococcus aureus (GRSA), and methicillin-resistant
Staphylococcus aureus (MRSA). Antibiotic-resistant bac-
teria, which are difficult to treat, have been reported to be
inactivated by using a nanosecond pulsed plasma or large-
area DBD-NBP plasma. [45].
As shown in Fig. 23a and b, the expression levels of
MRSA resistance genes ecA, mecI, mecRI, and femA
before and after plasma treatment are compared using
the Q-PCR (quantitative real time polymerase chain reac-
tion). The expressions of all resistance genes are known
to be remarkably reduced, which is related to changes in
the composition of the cell wall. The genes of multidrug
resistant bacteria (PRSA, MRSA and GRSA) are signifi-
cantly reduced after 20 min of plasma treatment, allowing
the superbacteria to be controlled very nicely. At this time,
peptidoglycan is revealed to have been oxidized by affect-
ing the molecular bonds of carbonyl groups in the bacterial
cell wall due to the synergistic action of reactive oxygen
and pH [45]. As shown in Fig. 24a and b, when a nano-
second pulsed plasma with an energy per pulse of 0.1 J
or less is used, the shock wave is revealed to play a major
role in deactivating bacteria. Its detailed mechanisms can
be revealed through studies of the interactions between
plasmas and liquids.

Fig. 23  a Gene expression analyses after NPP and Ar-DBD-NBP treatment: a wild type S. aureus, b PRSA, c MRSA, and d GRSA [45]. b
Experimental composition of bacteria with or without plasma treatment, as obtained using XPS [45]

Fig. 24  [Top: c] Colony of S.

aureus: (i) wild type, (ii) after
NPP treatment; (iii) wild type,
(iv) after Ar-DBD-NBP treat-
ment [45]. [bottom: d] SEM
images of the S. aureus: (i) wild
type, (ii) after NPP, and (iii)
after Ar-DBD-NBP treatment
[45]

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Plasma bioscience for medicine, agriculture and hygiene applications 837
Fig. 25  a Anti-tumor effect of silymarin nanoemulsion (SN) and
plasma activated PBS treatment in a mice model. a Changes the
tumor volume in xenograft mice models. b macroscopic observation
of a control group and dual treatment of NBP-activated PBS and SN
6.3 Apoptosis of intractable cancer cells
and healing of degenerative brain diseases
Plasmas can be used to cure intractable cancer lesions
through changes in DNA, apoptosis and cell cycle control
proteins through the interaction of NBP or nanoparticles
with them. Figure 25a shows the results of simultaneous
treatment of a silymarin nanoemulsion (SN) and phosphate
buffered saline (PBS) treated with air DBD-NBP in xeno-
grafted mice with G361 skin cancer cells. Reactive oxygen
radicals in cancer cells are increased by about 3 times
compared to cells without plasma treatment, and active
nitrogen is increased by about 2.5 times, which damages
the DNA of cancer cells and reduces the size of the tumor.
[132] In addition, malignant proteins, such as plaque and
tau knots of beta amyloid (Aβ) protein, and the Lewis body,
a variant of alpha-synuclein, will accumulate in neurons in
the cerebral cortex, which may become a normal protein
again by promoting immune activation and enhancing the
antioxidant function of nerve cells through the interac-
tion of plasma reactive species. In particular, when atomic
oxygen (O) in the reactive species existing in air, nitrogen,
and oxygen NBP plasma are used their neurons are dif-
ferentiated as shown in Fig. 25b, which is applicable to
degenerative brain diseases such as Alzheimer's disease
(dementia) and Parkinson's disease [20].
In addition, breast cancer cells that have recently devel-
oped resistance to anticancer drugs involve an epiimmune
mechanism with active oxygen of DBD-NBP, and the sen-
sitivity of anticancer drugs has been stably restored so that
and anticancer drugs can be used continuously. In addi-
tion, the plasma itself can target cancer tissues by restoring
and promoting the immune system [133].
to nude mice group bearing subcutaneous tumors on the right hind
flank and c tumor sizes [132]. b Representative SEM images of
mouse neural differentiation by air, ­N2, and ­O2 NBP synapses formed
in response to a 1 min exposure of NBP (scale bar = 50 um) [20]
6.4 Plasma application in cancer treatment
approach
In the last several years, non-thermal plasmas have been
found to be highly effective tools for the delivering reac-
tive species to cancer cells in vivo or in vitro. Interesting
initially the NTP was not designed to promote cancer cell
death by inducing oxidative stress-mediated cell death
through reactive species generation. However, over time,
developments in plasma diagnostics and technology meth-
ods have provided targeted delivery of controlled mixtures
of various reactive species to tumor cells to regulate many
events critical to their malignant behavior and eventually
to stimulate preferential cancer cell apoptosis and metasta-
sis by taking advantage of short-lived and long-lived spe-
cies such as H­ 2O2 [134–136]. Plasmas have very low gas
temperatures, which make them suitable for direct use on
living tissues [137]. In mid-2019, the first clinical trial of
a plasma was approved by the FDA and was used to extend
the 2-year survival of a 33-year old patient in a late stage
pancreatic cancer [138]. Notwithstanding the increased
number of favorable reports, which shows the plasma
efficiency and selectivity against a wide variety of cancer
cells, [137, 139], the functions of different plasma-induced
effects in cancer-cell malignant transitions, including drug
sensitization remain largely unknown. Prominently, triple-
negative breast cancer cells lacking required receptors for
clinical purposes have increased endogenous ROS levels
and are therefore, more sensitive compared to receptor-
positive breast cancer cells, signifying the importance of
plasmas in sensitizing these cells more efficiently over
another breast-cancer subtype [140, 141]
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Fig. 26  Adaptive plasma. a Concept of an adaptive plasma. Cold
plasma treatment in vivo or in vitro will trigger differing responses
in cancer cells and normal cells. The signal based on these responses
will be measured, recorded, and analyzed by using a plasma system.
Keidar et al. suggested that the use of a plasma could
be efficient for targeted cancer cells treatment due to its
flexibility in managing the plasma-generated reactive spe-
cies chemistry, which eventually responses to control the
molecular events involved in cancer progression (Fig. 26)
[141]. Plasma are well known to promotes preferential
cancer cell killing in a dose-dependent fashion, i.e., from
cell growth arrest to necrosis/apoptosis [143, 144]. Also,
cancer cells were found to be more sensitive to endoge-
nous ­H2O2 rather than exogenous ­H2O2 over normal coun-
terparts, possibly due to the presence of a high catalase
concentration on their cell surface, as tumor cells have
a high local concentration of a catalase on their cell sur-
face which defends them against exogenous ­H2O2 [145,
146]. Singlet oxygen generated from a chemical reaction
between peroxynitrite and H ­ 2O2 activates a high number
of singlet oxygen in cancer cells which triggers intracel-
lular reactive-species mediated apoptosis signaling [147].
In addition, ozone, besides H ­ 2O2 and singlet oxygen has
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E. H. Choi et al.
Through the feedback system, the chemical composition will be mod-
ulated through changes in the power supply, the gas composition, and
the distance between the plasma source and the cells. b Plasma self-
adaptation-based self-organization and pattern formation [142]
been implicated as another key player in the underlying
mechanism of plasma-mediated cancer apoptosis [148].
Apart from apoptotic cell death, recently, plasmas have
been shown to be efficient for inducing immunogenic cell
death (ICD) in melanoma and lung cancer cells [149, 150]
in vitro and in colorectal cancers in vivo, [151] where short-
lived reactive species such as nitric oxide and hydroxyl radi-
cals are found to be active constituents [142]. This study
provides the first confirmation that plasmas have potential
for cancer immunotherapy which can be applied clinically.
­H2O2 is an essential player in oxidative-stress induced
apoptosis [152, 153], but not in ICD [18]. Recently, Kau-
shik et al. reported that the growth of cancer cells could be
inhibited when they are co-cultured with plasma-stimulated
macrophages via iNOS TNF-α release. Interestingly, plas-
mas do not reduce immune cell viability after exposure.
These findings reveal that plasma generated reactive species
could activate cytotoxic macrophages for cancer cell death
[154]. In an additional study, the authors found that plasma
Plasma bioscience for medicine, agriculture and hygiene applications 839
Fig. 27  Non-thermal plasma induced immune cell activation for ICD
induction [155]
Fig. 28  Overview of cellular response mechanisms following LTP
treatment [159]
exposure could successfully differentiate pro-inflammatory
macrophages, which supports anti-cancer immune responses
in resistant tumor cells (Fig. 27) [155]. Reports in the litera-
ture have indicated that plasma application may be helpful
targeting immunosuppressive cancers by modifying the pro-
tumor microenvironment.
Recently, a group of researchers showed the capability
of plasma treatment for necrosis induction by means of
primary fibroblast cultures isolated from human oral tissue
[156]. Moreover, a plasma can cause another form of can-
cer cell death, named senescence, by initiating a calcium
influx in skin cancer cells [157]. Several studies highlighted
the importance of dose given by the plasma to cancer cells
because when the level that healthy can cells tolerate is
exceeded, cells themselves can be killed too (Fig. 28) [158,
159]. Plasma can act as switch to control the fate of cancer
cells due to its dose-dependent characteristics under certain
circumstances which makes its use favorable as an anti-
cancer approach. In another way, plasma exposure which is
a cocktail of chemical reactive species, and physical effects
(heat and UV), can be utilized to determine the condition of
healthy cells that human encounter in their daily life, such
as normal tissue and transition events during carcinogenesis
similar to ionizing radiation used in medical field.
Usually, in advanced cancer stages, oncologists prefer
radiotherapy widely for malignant and invasive tumors;
however, normal tissue side-effects and developed resistance
reduce the success rate of radiotherapy in cancer patients.
Recently, plasma treatment has been revealed to restore
cell sensitivity to traditional therapeutic modalities, such
as temozolomide and tamoxifen, in resistant gliomas [160]
and breast cancer [161] cells and to slow integrin signaling
pathways that facilitate chemo- and radio-resistance in dif-
ferent cancer cells (Fig. 29) [162]. Instead, researchers also
tried to improve plasma effect on cancer selectivity by tar-
geting cancer cell metabolism. In order, Kaushik et al. used
the combination approach of plasma and 2-Deoxy d-glu-
cose (2-DG) to destroy blood cancer cells, where effective
anti-cancer effects were achieved at low doses of plasma.
This study claims that the addition of 2-DG enhances the
plasma efficiency by targeting the cancer glycolysis pathway,
which eventually leads to cancer cell apoptosis by intrin-
sic pathways [11]. They further extended their study and
tested the effect of silymarin nano-emulsion and plasma
co-treatment on skin cancer cells. Remarkably, this dual-
treatment approach impaired the epithelial-mesenchymal
transition (EMT) and cancer stem cell maintenance observed
as in vitro and in vivo [132]. Cancer cells believed to be are
extremely heterogeneous, which makes them susceptible or
resistant to particular treatments, including radiotherapy and
chemotherapy. Since plasma generates a mixtures of various
chemical species and particles, it can target different cellular
biomolecules or associated pathways in this heterogeneous
cancer cell population. This property of a plasma makes it
potentially more effective over traditional methods.
Moreover, plasma selectivity can be achieved by using
functionalized nanomaterials having specific target recog-
nition in cancer cells [163]. Using this concept, to analyze
plasma activity induced using nanomaterials, Kaushik et al.
investigated the effects of a combination of PEG-coated gold
nanoparticles and a plasma on the tumor metastasis. It is
claimed that cancer cell growth was inhibited by deactiva-
tion of the PI3K/AKT signaling pathway, along with reversal
of EMT, as was observed by a delay in tumor growth in
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Fig. 29  Treatment of tamoxifen resistant MCF7 breast cancer cells by using plasma and gene expression analysis to uncover mechanism related
to for plasma-induced cancer cell sensitization [161]
a tumor xenograft mice models [66]. Having a successful
plasma approach in nanoparticle formulation, Linh et al.
fabricated polydopamine-functionalized gold nanoparticles,
avoiding the use of added toxic chemicals. Specially, plasma
treatment reduced the reaction time essential for the syn-
thesis of these particle unlike traditional chemical methods.
Worth mentioning is that these synthesized have increased
cellular uptake and cytotoxic effect, as seen in breast cancer
cells, which emphasize the significance of plasma–fabricated
PDA-GNPs for inhibition of cancer cell growth through tar-
geted delivery inside the cells [164].
For cancer treatment, plasmas can be generated using
various plasma sources or alternatively can be administered
indirectly to cancer cells in the form of a liquid such as a
plasma-activated medium (PAM), making it a quite flexible
tool for clinical purposes [165]. In direct plasma treatment,
only confined treatments are achievable. Nonetheless, the
PAM selectivity against cancers is subjective, such as cell
type as compared to direct plasma treatment [166, 167].
Many scientists have proposed that cancer-cell death induced
by PAM largely relies on the combination of N ­ O2- and H
­ 2O2
[168, 169]. Given the short incubation times of plasma-oxi-
dized solutions, such as Ringer's lactate, sodium chloride,
and PBS, such solutions can be beneficial in improving the
efficacy of in vitro experiments [170, 171]. In another study,
repeated treatment with PAM by intraperitoneal injection
was found to reduce the mice tumor growth as seen by MRI,
leading to improved survival rate. This group suggested
that due to the nominal side effects of such plasma-oxidized
solutions, they are suitable therapeutic tools for treating
advanced stages tumor [171]. However, further conclusions
are definitely needed to determine whether PAM treatment
can prevent tumor relapse by using xenograft mouse mod-
els [74]. For clinical safety assessment of plasma, Isbary
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E. H. Choi et al.
et al. conducted ex vivo experiments using human skin tis-
sues, where a 2 min plasma treatment did not induce DNA
damage to human cells [172]. Currently, clinical testing of
plasma safety and efficiency is being considered regularly
in treatments of chronic cutaneous ulcers in phase I or II tri-
als. Plasma treatment was feasibly tolerated in most cases,
without any frequent adverse effect [173]. Our primary per-
ceptions are not restricted to only cancer research using plas-
mas. Plasma can also be advantageous for the treating other
deadly human diseases such as neurological disorders [174].
6.5 Cellular mechanism of NBP as plasma treated
liquid for cancer
NBP has been reported to have a selective killing effect on
targeted cancer cells with minimal effects on regular cells
that act by generating reactive oxygen species (ROS) and
reactive nitrogen species (RNS). In previous study, we
assessed the effects of a non-thermal air soft jet plasma on
the U87 brain cancer by using the physicochemical and bio-
logical (PCB) correlation between the RONS cascade and
MAPK/PI3K/AKT signaling pathway, which resulted in
apoptosis [175, 176]. The results showed that the soft jet
plasma significantly inhibited cell proliferation, induced cell
cycle arrest, inhibited survival pathway and induced apop-
tosis pathway in U87 cells (Fig. 30).
Both gaseous and liquids plasmas are acts on this con-
cept. It is possible to capture the plasma particles by gen-
erated gas type of plasma into liquid. Comparison of the
features of NBP and NO water as a plasma-treated liquid
(PTL) for cervical cancer cells [177]. PTL was reported
to have a cytotoxicity for various cancer cells like NBP,
and both use the same cellular mechanisms to induce
DNA damage and apoptosis. We have been developed a
Plasma bioscience for medicine, agriculture and hygiene applications 841
Fig. 30  Diagram of cellular
mechanism underlying NBP in
brain tumor
microwave jet plasma system producing nitric oxide for
aqueous soluble NO (nitric oxide) in distilled water at
room temperature. The concentration of nitric oxide could
controlled by using the gas flow and the microwave power.
According this results, we sought to explore the effects on
cervical cancer cells of NO water as PTL on cervical can-
cer cell line compare with DBD-NBP. Therefore, although
some technical challenges remain to be addressed, unlike
radiation, which has been thoroughly studied, DBD-NBP
and NO-PAM have the potential to be new application for
effective and safe clinical trials (Fig. 31).
In-vivo studies have revealed that NBP-PTW showed
anticancer effects on chemo-resistant lung cancers, and on
non-small cell lung cancer (NSCLC). The effect were simi-
lar to these aspect of a cancer drug, gefitinib drug, in the
chemo-resistant NSCLC model [178]. Gefitinib is a epider-
mal growth factor receptor-tyrosine kinase inhibitor (EGFR-
TKIs) and has been shown to have significant benefits for
treating NSCLC. The anticancer activities of PTW seems to
be involved in inhibiting proliferation and in angiogenesis
and enhancing apoptosis in tumor cells (Fig. 32). Interest-
ingly, PTW contributes to an enhanced immune response
and improved cachexia in the model.
The NBP as a gas stage and a liquid stage is shown to
have systemic application (oral administration) and apical
application (direct administration) in the treatment of cancer
cells subsequently it can be useful tool for clinical trial to
improve of cachexia and immunostimulation.
6.6 Regenerative medicine for stem cell
modification by using the NBP
The NBP can use tissue engineering to stimulate stem cells
and scaffold [179]. When discharging the NBP to stem cells,
differentiation of stem cells without other stimuli induced by
bone cells has been reported. The NBP generates fast ions,
and free radicals and then immediately reacts with surface of
molecules subsequently occur the oxidation/reduction reac-
tion. In stem cells, can induce differentiation by reactive
species by providing various stimuli to the cells.
Also, the NBP can stimulate to osteogenic initiation with-
out osteogenic induction molecules like ascorbic acid on
MC3T3-E1 cells [180]. MC3T3-E1 is an osteoblastic precur-
sor cell line from C57BL/6 mouse calvaria tissue. MC3T3-
E1 has stimulated the osteogenic differentiation when treated
with ascorbic acid and beta-glycerophosphate to forming
osteogenic molecules such as alkaline phosphatase (ALP)
osteocalcin (OCN), osteopontin (OSP), collagen type 1
(COL1), and Runx2.
We hypothesized that the non-thermal plasma can stimu-
lated osteogenic initiation without osteogenic induction mol-
ecules like ascorbic acid. Here we claim the effect on the
osteogenic potential of NBP on stem cells, such as human
bone marrow stem cells (hBMSCs) and human periodontal
ligament stem cells (hPDLSCs) [181]. The stem cells were
cultured and then treated with NBP with or without induc-
tion medium. We evaluated the differentiation rate of cells
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Fig. 31  Apoptosis of HeLa cells after NBP and NO-PAM treatment.
HeLa cells were treated with NBP for different times and with NO-
PAM different concentrations. Dot plots in a and histograms in b for
NBP (upper) and NO-PAM (lower) showed that both longer-time
NBP treatment and higher-concentration NO-PAM treatment could
by using real time PCR. It increased BMSCs and PDLSCs.
Nonthermal plasma treatment without osteogenic induc-
tion medium. The osteogenic differentiation marker, OCN,
osterix, Runx2 and ALP expression level was increased in
those cells. PDLSC cells were treated with a plasma for dif-
ferent time and then characterized by real time PCR. As
depends on treated time, it was clearly observed increasing
the OCN, Runx2, COL1 by the plasma treatment.
Our study demonstrated that NBP has the ability to
induce osteogenic differentiation of hMSCs. Moreover, we
found that MSCs from different tissue sources had different
responses to NBP-induced osteogenesis, while hPDLSCs
had more compliance and controllability than hBMSCs.
More importantly, we have deeply studied the reason for the
effect of NBP, and we believe that NBP treatment activates
ROS-sensitive molecules, p38 MAPK, and that the acti-
vated p38 MAPK further stimulates antioxidants for cellular
homeostasis and FoxO1 expression to modulate osteogenic
differentiation in hPDLSCs (Fig. 33). Our study reveals for
the first time the mechanisms underlying NBP and hPDLSC,
osteogenic differentiation, which lays the foundation for the
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E. H. Choi et al.
induce more Hela cell apoptosis, compared with control group. c
Western Blots performed to confirm the apoptosis effect of NO-PAM
inducing apoptosis in Hela cells with NO concentration of 7 μM, and
incubated for 1, 2, 4, and 6 h after treatment
application of NBP in bone tissue regeneration and engi-
neering the NBP can induce new bone. In additional, it
may be use combinations of conventional therapeutic treat-
ment, as well as new approaches, to target tissue for bone
regeneration.
6.7 Plasma application to COVID‑19 (SARS‑COV2:
Coronavirus 19)
SARS-COV-2, or COVID-19, originated in 2019, has now
spread worldwide at an alarming rate, thus infected about
several million people, and has caused about millions of
deaths, with an approximate death rate of 2.2%. In addition,
confirmed cases continue to spread infected in Asia, US,
Europe, Africa and all around the world. Now, many kinds
of vaccines against COVID-19 have been developed, and
some of countries are now vaccinating people to achieve
herd immunity. Viruses play an important role in the global
ecological environment, such as the carbon cycle of the sea
[182]. On the other hand, pathogenic viruses infect tens of
millions to hundreds of millions of animals each year on the
Plasma bioscience for medicine, agriculture and hygiene applications 843

Fig. 32  Immunohistochemistry in tumor mass. Tumor tissue was enal (4HNE) for oxidative stress, and iNOS, TNF-α for immune
stained for caspase-3, PARP for apoptosis, Ki-67 for cell prolifera- response (A). The percentages of immune-reactive cells are shown in
tion, CD31 for angiogenesis, nitrotyrosine (NT) and 4-hydroxynon- (B–E) [178]

Fig. 33  Schematic diagram for

plasma effects on precursor
cells for osteogenic differentia-
tion through FOXO pathway
stimulated by p38 signaling

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844 E. H. Choi et al.

planet or infect people. This has resulted in a decrease in

crop yield and a loss of many lives. Therefore, inactivation
of harmful viruses is an essential and very important issue
for human health.
In the last 2-decade, three coronavirus (CoV) outbreaks
(SARS 2002, MERS 2013, SARS-COV-2 2019) have
occurred. SARS-CoV-2 (COVID-19), a major threat to
human health and the world economy, has a high homol-
ogy of 80% with SARS-CoV and 50% with MERS-CoV,
all of which are beta-coronaviruses. Therefore, the future
threat virus (named CoV-X) likely be a variant of it. Despite
repetitive pandemics such as SARS-CoV, MERS-CoV,
SARS-CoV-2, etc., we do not have date from of a preemp-
tive comparative analysis of the virological, immunologi-
cal and pathological characteristics of the strains or viruses
that have not yet spread from animals to humans. With the
proposition that the coronavirus continues to repeat the pan-
demic of human infection, the follow-up work of emergency
investigations on SARS-CoV-2 might be a potential help to
find risk factors and treatment options for a future pandemic.
Preemptive investigations on plasma treatment of coronavi-
rus strains or other host-derived species are needed in future. Fig. 34  Inactivation of viruses using a nonthermal biocompat-
SARS-COV2, which originated in Wuhan, China in ible plasma. A Various types of viruses (morphologically different)
treated with nonthermal plasma. B Magnified plasma properties
December 2019, has now spread globally at an alarming responsible for virus inactivation. ROS, RNS, UV radiation, and ions
rate and other variants of concerns are emerging after every and electrons (charged particles) are the most essential constituents
few months, still infecting about million of people, and caus- for virus inactivation. A NBP can target both viral nucleic acid and
ing about several deaths by an approximate death rate of proteins. C After NBP treatment, the virus proteins, and the genetic
materials such as DNA, RNA are degraded to non-infective particles
more than 2%. In addition, variant positive cases continue that cannot affect its hosts. [44]
to spread in the US, Australia, Brazil, Europe, Africa, Asia
and other part of world. Viruses have been dominate for the
over billions of years on the earth [182], and many types of strains, which are highly pathogenic, can also be inactivated
them exist and have adapted, evolved, and mutated according by using a nonthermal plasma, which is considered a new
to the situations. hope for virus treatments [44]. This virus-based research
Viruses are important microbial predators that influence area is still relatively young and started about two decade
global biogeochemical cycles and drive microbial evolution, ago, almost coincident with the perspective of plasma bio-
although their impact is often under appreciated. Viruses science researches [183–185].
play an essential role in the carbon cycle of the sea [182]. On In the early 90 s, ozone was known to be able to inactivate
the other hand, pathogenic viruses infect millions of human the virus. Due to many safety issues for using only ozone
and animals every year on the earth. Thus, the inactivation gas, many groups are conducting their investigations on the
or suppression or control of pathogenic viruses is a critical inactivation of viruses by using other eco-friendly ionized
and important issue for the health of humans and animals. gases while maintaining ozone some level. NBP plasmas
The inactivation or suppression of a virus can be success- have been commonly used in several areas for microbial
fully processed using reactive oxygen and reactive nitrogen inactivation or decontamination. Recently, a few plasma
in a plasma [183]. Figure 34 shows a plasma consisting of research groups initiated preliminary investigations on
electrons, ions, and cocktail of reactive oxygens and nitrogen corona viruses by using various kinds of plasma devices and
species at room temperature, that can inactivate various type treatment procedures [44, 46, 48, 49, 186–188]. Recently,
of viruses. The reactive species cocktail interaction with viruses were shown to be using plasma exposure and plasma
viruses causes damages to genetic material such as DNA or activation liquids such as water, media, buffers, clinical solu-
RNA and proteins of the viruses, as shown in Fig. 34. In the tions etc. Figure 35 shows the effectiveness of plasma jets in
case of nucleic acid, a plasma can damage DNA and RNA, removing corona viruses from the surfaces of objects [187].
can reduce cell replication, and can inhibit the proliferation Both plasma activated water and plasma treatment showed
of viruses due to the enzyme activity caused by oxidation inactivation of viruses in an exposure dose dependent man-
of lipids and proteins. SARS-COV-2 virus and its variant ner, and investigations suggested no side-effects with no

13 Vol.:(0123456789)
Plasma bioscience for medicine, agriculture and hygiene applications 845
Fig. 35  Argon gas-based plasma discharge disinfecting SARS-
CoV-2: a Argon plasma treatment of a plastic surface and the opti-
cal emission spectra of ROS and RNS (exposure: 250 ms), and b
the bright-field image of Vero-E6 cells infected with SARS-CoV-2
showing the viral cytopathic effect (CPE). Control of uninfected cells
or minimal invasiveness, along with ease of use, enhanced
wound healing, and inhibition of inflammation [44, 188].
Recently Guo et al. investigated plasma activated water
(PAW) as an alternative disinfectant [49]. Disruption of
the SARS-COV-2 infection process by damaging the spike
protein (S-protein) of the receptor-binding domain (RBD)
and by binding to the cellular receptor human angiotensin-
converting enzyme 2 (hACE2) was the main target of that
study. In the study, pseudoviruses with the S protein of
SARS-CoV-2 were used as a model and related mechanisms.
The PAW was prepared using a surface discharge plasma
device consisting a water-facing grounded mesh electrode
and high-voltage plane electrode separated by a polytetra-
fluoroethylene dielectric layer (Fig. 36). Their aim was to
check the antiviral ability of PAW against pseudovirus, thus
providing a new disinfection method to fight the pandemic.
The PAW damaged or inactivated S-protein incorporated in
was included in the experiment. c Titre response on SARS-CoV-2
to plasma treatment exposures of 0, 30, 60, and 180 s on surfaces of
metal, cardboard, plastic, football leather, composite, baseball leather
and basketball leather. The distance between the object surface and
the plasma device was ∼15 mm [35]
the pseudovirus of SAR-COV-2 and inhibited the infection
process. Species such as ­ONOO– and ­O2•–, (Short-lived spe-
cies generated in PAW) are assumed to react selectively with
proteins. This study will be a nice example for using PAW
as a disinfection agent due to its effectiveness, even after
long storage (more than 30 days), and strength compared to
market available hydrogen peroxide. Thus long-term effec-
tiveness of PAW was shown to provide a basis for the devel-
opment of a new disinfectant. Further studies on the role of
PAW as a disinfectant for SARS-COV-2 and other corona
viruses may further support this investigation.
Several investigations have described the effect of cold
plasma treatment on various viruses, such as bacteriophages
(MS2, Φ174, and T4 strains), human norovirus (NV), feline
calicivirus (FCV), porcine reproductive and respiratory syn-
drome (PPRS), Newcastle disease virus (NDV), murine nor-
ovirus (MNV), respiratory syncytial virus (RSV), hepatitis
13 Vol.:(0123456789)
846
Fig. 36  Plasma-treated water (PAW) as a disinfection agent. a Schematic diagram of the pseudovirus study and b PAW preparation method
using a surface plasma device [49]
A virus (HAV), etc. Plasma-induced ROS and RNS, ozone,
as well as some amount of UV, electrons, ions, and other
factors can affect structural components such as cell walls
or membranes, along with physiological components such as
nuclear material and proteins. Nonetheless, definite evidence
for the inactivation of virus pathogenesis and a modification
of the host–pathogen interaction mechanism by using plasma
has been not reported yet. The necessity to study plasma-
based virus deactivation mechanisms in both animal models,
and cell-based models including organoids against future
variants of corona viruses is increasing. The mechanism for
the inactivation and control of Coronaviruses will aim at
host–pathogen interaction and virulence factor that can pro-
vide new perceptiveness for the control of virus pathogenesis
by using plasmas.
Human coronaviruses, such as HCoV-229E and HCoV-
OC43, HCoV-NL63 and HCoV-HKU1, usually cause
mild respiratory tract infections whereas severe acute
respiratory syndrome coronavirus (SARS-CoV), Middle
East respiratory syndrome coronavirus (MERS-CoV) and
SARS-CoV-2, are highly pathogenic. These viruses are
present in intermediate hosts such as bats wherein they
undergo recombination and mutations forming altered
13 Vol.:(0123456789)
E. H. Choi et al.
genotypes leading to severe pathogenicity. SARS-CoV-2
enters into human cells by using an interaction between
receptor-binding domain (RBD) of SARS-CoV-2 spike
(S) protein and the cell surface receptor angiotensin-con-
verting enzyme-2 (ACE2). The viral S-protein further
undergoes cleavage by proteases (TMPRS2) or cathepsin
B or FURIN to cause fusion of the host cell membrane
and an ensuing infection. Recently, studies have shown
that the SARS-CoV-2-specific virulence factors are pro-
grammed to evade detection and to modulate host immune
cells responses, including interferon (IFN) regulation and
to utilize the host cell for viral replication. Although sev-
eral studies have established the role of Spike protein
as a virulence factor of SARS-COV-2, the recent variant
of SARS-Cov-2 (deletion 69–70, deletion 144, N501Y,
A570D, D614G, P681H, T716I, S982A, D1118H) has
created open questions as to our understanding of vir-
ulence factors. Speculation is that more mutations and
variants might emerge from the coronavirus family and
might cause future corona virus epidemics or pandem-
ics. A possibility exists that every time we may have to
design new vaccines for future corona virus strains. This
makes in-depth investigations for plasma-based treatment
Plasma bioscience for medicine, agriculture and hygiene applications 847
procedures to control functions of virulence factors or to
damage viral genetic material essential.
Plasma-based future research must be based on topics
related to use of model viruses (including low pathogenic
coronavirus) that can quickly respond to future corona
virus threats. To attain this aim, the plasma community
must understand the general virological characteristics of
coronavirus: invasion into cells during infection, prolif-
eration within cells, release of viruses after proliferation,
and transfer to adjacent cells. Finally, we can analyze
important factors of pathogenicity after plasma treat-
ment in the coronavirus infection process build advance
information, and secure plasma treatment technology for
future viruses. For this purpose, we need to focus on viro-
logical factors (such as coronavirus infection, replication,
life cycle research), immunological factors (such as host
immune interactions and immune specificity), pathologi-
cal factors (research on biological pathogenesis and path-
ogenicity), and infection characteristics of mutant strains
and animal viruses. Using these parameters for analysis
after plasma treatment, we can establish a next-generation
pandemic infectious disease treatment and sterilization
system.
Also, artificial intelligence (AI)-based prediction mod-
els can be used to predict possible treatment strategies for
the next upcoming mutated virus strains. These AI mod-
els can assist in early prediction of virulence properties,
which can help to support plasma-based treatments. The
development of cost-effective economical plasma-based
treatments will provide endless prospects for the treat-
ment of critical stage patients infected with the virus. Fur-
thermore, now is the time to perform investigations and
to develop plasma-based devices or products for active
treatment of COVID-19 and its variant virus around the
world while ensuring safety of the patients and the medi-
cal doctors.
Due to environmental safety standards, many research-
ers are conducting their investigations on inactivation of
viruses by using other eco-friendly reactive gases while
avoiding high amount of ozone. In addition, the reality
is that research on the interaction between a plasma and
a virus needs to be recognized as important for clarify-
ing how to inactivate COVID-19 and its variant virus for
the development of plasma quarantine devices for human
beings. Furthermore, now is the time to conduct research
and development while ensuring the safety of patients and
medical doctors on the implementation of plasma treat-
ment devices for active treatment of COVID-19 and its
variant virus around a world. In addition, the simultaneous
development of plasma-treated water needs to be empha-
sized because the justifications for its use are plentiful.
7 Perspectives and conclusion
Plasma bioscience and medicine are newly developed
and rapidly developing, with the goal of next-generation
health promotion and disease healing. To this end, the
development of nonthermal (or low temperature) atmos-
pheric pressure biocompatible plasma (NBP) source that
can be actually applied in the medical field, and research
and development of plasma activated water (PAW) using
this NBP are actively underway. These NBP and PTW
can also be applied to cancer cells, neurodegenerative dis-
eases, and skin diseases, which have difficult treatments.
Active cooperative research, such as securing new data
and related basic plasma clinical trials are needed. The
treatment based on plasma bioscience can be combined
with existing cancer treatment methods and can be applied
in various ways. For further exploration of recent research
has investigated the effect of plasma-treated biomolecules
for cancer treatment application [189]. These kind of
plasma-treated or activated biomolecules also have the
potential for numerous biomedical applications. In par-
ticular, the NBP can be expected to have great effects when
it can be applied together with plasma treated water to
treat liver cancer, lung cancer, brain cancer, breast cancer,
pancreatic cancer, skin cancer, and blood cancer, which
are known to be types of intractable cancer. Nonthermal
plasma-synthesized nanoparticles can also be utilized for
cancer treatment and several other biomedical applica-
tions. These environmentally friendly synthesized nano-
particles can induce immunogenic cell death and release
a damage-associated molecular pattern in cancer cells.
Moreover, these nanoparticles exhibit enhanced uptake in
malignant cells and less toxicity in normal cells [190]. In
addition, the manufacture of, and disinfection of plasma
medical devices for surgical trauma treatment, wound
healing, burn treatment, and skin disease control and treat-
ment are recognized as very important fields. Moreover,
the NBP can be expected to be efficient when sterilizing
bacteria resistant to antibiotics. Recent investigations in
plasma bioscience and medicine showed the application
of a gas plasma for bio-sterilization, decontamination, and
water purification [191, 192]. Also, these plasma sources
or plasma-activated water with minerals can be used for
agriculture applications [193], and this area can be fur-
ther explored with several new plasma-based sustainable
agriculture ideas. Now, in the era of long-lived health and
welfare, it is also important to securing cooperative tech-
nology to solve this by applying NBP to degenerative brain
diseases among the silver generation is important.
In 2015, we need to get a database of realistic wisdom
about the valuable experience with the Middle East Res-
piratory Syndrome (MERS) and the infection of the new
13 Vol.:(0123456789)
848
virus COVID-19 this year, as seen in the harsh reality.
Prevention and inactivation of these viruses in all-weather
areas, indoor disinfection of ambulances, indoor air purifi-
cation and virus removal, water purification and fine dust
removal are difficult to implement only with conventional
methods. At this time, applying NBP technology can bring
better results than expected.
Acknowledgements This study was supported by the National
Research Foundation (NRF) of Korea, funded by the Korea govern-
ment (NRF-2016K1A4A3914113, 2021R1A6A1A03038785).
References
1. F. Massines, G. Gouda, J. Phys. D. Appl. Phys. 31, 3411–3420
(1998)
2. U. Kogelschatz, Plasma Chem. Plasma Process. 23, 1–46 (2003)
3. A. Fridman, A. Chirokov, A. Gutsol, J. Phys. D. Appl. Phys. 38,
R1–R24 (2005)
4. J.S. Sousa, G. Bauville, B. Lacour, V. Puech, M. Touzeau, J.-L.
Ravanat, Appl. Phys. Lett. 97, 141502 (2010)
5. M.G. Kong, G. Kroesen, G. Morfill, T. Nosenko, T. Shimizu, J.
Van Dijk, J.L. Zimmermann, New J. Phys. 11, 115012 (2009)
6. J. Winter, K. Wende, K. Masur, S. Iseni, M. Dünnbier, M.U.
Hammer, H. Tresp, K.D. Weltmann, S. Reuter, J. Phys. D. Appl.
Phys. 46, 295401 (2013)
7. P. Attri, Y.H. Kim, D.H. Park, J.H. Park, Y.J. Hong, H.S. Uhm,
K.-N. Kim, A. Fridman, E.H. Choi, Sci. Rep. 5, 1–8 (2015)
8. E.H. Choi, H.S. Uhm, N.K. Kaushik, AAPPS Bull. 31, 1–38
(2021)
9. N. Barekzi, M. Laroussi, IEEE Trans. Plasma Sci. 42, 2738–2739
(2014)
10. N. Barekzi, M. Laroussi, Plasma Process. Polym. 10, 1039–1050
(2013)
11. N. Kaushik, N. Uddin, G.B. Sim, Y.J. Hong, K.Y. Baik, C.H.
Kim, S.J. Lee, N.K. Kaushik, E.H. Choi, Sci. Rep. 5, 1–11 (2015)
12. E.H. Choi, Vac. Mag. 7, 2 (2020)
13. K.N.K. E. H. Choi, Y. H. Kim, Vac. Mag. 38 (2014)
14. E.H. Choi, Vac. Mag. 9 (2015)
15. G.L. Geison, Isis 60, 273–292 (1969)
16. S.L. Miller et al., Science 117, 528–529 (1953)
17. J.D. Watson, F.H.C. Crick, Nature 171, 737–738 (1953)
18. T. von Woedtke, A. Schmidt, S. Bekeschus, K. Wende, K.-D.
Weltmann, In Vivo (Brooklyn) 33, 1011–1026 (2019)
19. M. Laroussi, Front. Phys. 8, 74 (2020)
20. J.-Y. Jang, Y.J. Hong, J. Lim, J.S. Choi, E.H. Choi, S. Kang, H.
Rhim, Biomaterials 156, 258–273 (2018)
21. X. Lu, M. Keidar, M. Laroussi, E. Choi, E.J. Szili, K. Ostrikov,
Mater. Sci. Eng. R Rep. 138, 36–59 (2019)
22. D. Dobrynin, G. Fridman, G. Friedman, A. Fridman, New J.
Phys. 11, 115020 (2009)
23. M. Ito, J.-S. Oh, T. Ohta, M. Shiratani, M. Hori, Plasma Process.
Polym. 15, 1700073 (2018)
24. J.-S. Song, S.B. Kim, S. Ryu, J. Oh, D.-S. Kim, Front. Plant Sci.
11, 988 (2020)
25. C. Smet, M. Baka, A. Dickenson, J.L. Walsh, V.P. Valdramidis,
J.F. Van Impe, Plasma Process. Polym. 15, 1700048 (2018)
26. B. Adhikari, M. Adhikari, B. Ghimire, G. Park, E.H. Choi, Sci.
Rep. 9, 1–15 (2019)
27. P. Attri, K. Ishikawa, T. Okumura, K. Koga, M. Shiratani, Pro-
cesses 8, 1002 (2020)
13 Vol.:(0123456789)
E. H. Choi et al.
28. B. Adhikari, K. Pangomm, M. Veerana, S. Mitra, G. Park, Front.
Plant Sci. 11, 77 (2020)
29. P.J. Cullen, J. Lalor, L. Scally, D. Boehm, V. Milosavljević, P.
Bourke, K. Keener, Plasma Process. Polym. 15, 1700085 (2018)
30. G. Pauzaite, A. Malakauskiene, Z. Nauciene, R. Zukiene, I. Fila-
tova, V. Lyushkevich, I. Azarko, V. Mildaziene, Plasma Process.
Polym. 15, 1700068 (2018)
31. V. Mildaziene, G. Pauzaite, Z. Naucienė, A. Malakauskiene, R.
Zukiene, I. Januskaitiene, V. Jakstas, L. Ivanauskas, I. Filatova,
V. Lyushkevich, Plasma Process. Polym. 15, 1700059 (2018)
32. C. Szőke, Z. Nagy, K. Gierczik, A. Székely, T. Spitkól, Z.T.
Zsuboril, G. Galiba, C.L. Marton, K. Kutasi, Plasma Process.
Polym. 15, 1700138 (2018)
33. Y. Park, K.S. Oh, J. Oh, D.C. Seok, S.B. Kim, S.J. Yoo, M.-J.
Lee, Plasma Process. Polym. 15, 1600056 (2018)
34. V. Štěpánová, P. Slavíček, J. Kelar, J. Prášil, M. Smékal, M. Stu-
pavská, J. Jurmanová, M. Černák, Plasma Process. Polym. 15,
1700076 (2018)
35. J.-S. Kwon, S.-H. Choi, E.H. Choi, K.-M. Kim, P.K. Chu, Int. J.
Mol. Sci. 21, 6085 (2020)
36. M.-J. Lee, J.-S. Kwon, H.B. Jiang, E.H. Choi, G. Park, K.-M.
Kim, Sci. Rep. 9, 1–13 (2019)
37. S.-H. Seo, I. Han, H.S. Lee, J.J. Choi, E.H. Choi, K.-N. Kim, G.
Park, K.-M. Kim, Sci. Rep. 7, 1–11 (2017)
38. T.T. Gupta, S.B. Karki, J.S. Matson, D.J. Gehling, H. Ayan,
Biomed Res. Int. 2017, 6085741 (2017)
39. H. Wu, C. Moser, H.-Z. Wang, N. Høiby, Z.-J. Song, Int. J. Oral
Sci. 7, 1–7 (2015)
40. B. Gomes, S.F.C. Souza, C.C.R. Ferraz, F.B. Teixeira, A.A. Zaia,
L. Valdrighi, F.J. Souza-Filho et al., Int. Endod. J. 36, 267–275
(2003)
41. S.R. Herbst, M. Hertel, H. Ballout, P. Pierdzioch, K.-D. Welt-
mann, H.C. Wirtz, S. Abu-Sirhan, E. Kostka, S. Paris, S. Preiss-
ner, Open Dent. J. 9, 486 (2015)
42. R.O. Darouiche, N. Engl, J. Med. 350, 1422–1429 (2004)
43. L. Hall-Stoodley, J.W. Costerton, P. Stoodley, Nat. Rev. Micro-
biol. 2, 95–108 (2004)
44. A. Filipić, I. Gutierrez-Aguirre, G. Primc, M. Mozetič, D. Dob-
nik, Trends Biotechnol. 38(11), 1278–1291 (2020)
45. J.H. Park, N. Kumar, D.H. Park, M. Yusupov, E.C. Neyts, C.C.W.
Verlackt, A. Bogaerts, M.H. Kang, H.S. Uhm, E.H. Choi et al.,
Sci. Rep. 5, 1–14 (2015)
46. R. Kar, N. Chand, A. Bute, N. Maiti, A.N. Rao, V. Nagar, R.
Shashidhar, D.S. Patil, S.K. Ghosh, A. Sharma, Trans. Indian
Natl. Acad. Eng. 5, 327–331 (2020)
47. A. Bisag, P. Isabelli, R. Laurita, C. Bucci, F. Capelli, G. Dirani,
M. Gherardi, G. Laghi, A. Paglianti, V. Sambri, V. Colombo,
Plasma Process. Polym. 17, 2000154 (2020)
48. T. Xia, A. Kleinheksel, E.M. Lee, Z. Qiao, K.R. Wigginton, H.L.
Clack, J. Phys. D. Appl. Phys. 52, 255201 (2019)
49. L. Guo, Z. Yao, L. Yang, H. Zhang, Y. Qi, L. Gou, W. Xi, D. Liu,
L. Zhang, Y. Cheng, X. Wang, M. Rong, H. Chen, M.G. Kong,
Chem. Eng. J. 421, 127742 (2020)
50. T. Bernhardt, M.L. Semmler, M. Schäfer, S. Bekeschus, S.
Emmert, L. Boeckmann, Oxid. Med. Cell. Longev. 2019, 1–10
(2019)
51. T. von Woedtke, S. Emmert, H.-R. Metelmann, S. Rupf, K.-D.
Weltmann, Phys. Plasmas 27, 70601 (2020)
52. N.K. Kaushik, N. Kaushik, R. Wahab, P. Bhartiya, N.N. Linh, F.
Khan, A.A. Al-Khedhairy, E.H. Choi, Cancers (Basel) 12, 457
(2020)
53. A.A. Fridman, G.G. Friedman, Plasma medicine (Wiley, Chich-
ester, 2013)
54. M. Laroussi, M.G. Kong, G. Morfill, W. Stolz, Plasma medicine:
applications of low-temperature gas plasmas in medicine and
biology (Cambridge University Press, Cambridge, 2012)
Plasma bioscience for medicine, agriculture and hygiene applications 849
55. A. Bogaerts, M. Yusupov, J. der Paal, C.C.W. Verlackt, E.C.
Neyts, Plasma Process. Polym. 11, 1156–1168 (2014)
56. H. Tanaka, M. Hori, J. Clin. Biochem. Nutr. 60, 16–67 (2017)
57. H.-R. Metelmann, T. Von Woedtke, K.-D. Weltmann, Com-
prehensive clinical plasma medicine: cold physical plasma for
medical application (Springer, 2018)
58. S. Toyokuni, Y. Ikehara, F. Kikkawa, M. Hori, Plasma medical
science (Academic Press, 2018)
59. X. Lu, G.V. Naidis, M. Laroussi, S. Reuter, D.B. Graves, K.
Ostrikov, Phys. Rep. 630, 1–84 (2016)
60. E.J. Szili, J.-S. Oh, H. Fukuhara, R. Bhatia, N. Gaur, C.K.
Nguyen, S.-H. Hong, S. Ito, K. Ogawa, C. Kawada, Plasma
Sourc. Sci. Technol. 27, 14001 (2017)
61. D. Liu, E.J. Szili, K. Ken-Ostrikov, Plasma Process. Polym. 17,
2000097 (2020)
62. E. Turrini, A. Stancampiano, E. Simoncelli, R. Laurita, E. Cat-
anzaro, C. Calcabrini, M. Gherardi, V. Colombo, C. Fimognari,
Clin. Plasma Med. 9, 15–16 (2018)
63. N. Kaushik, S.J. Lee, T.G. Choi, K.Y. Baik, H.S. Uhm, C.H.
Kim, N.K. Kaushik, E.H. Choi, Sci. Rep. 5, 1–11 (2015)
64. I. Adamovich, S.D. Baalrud, A. Bogaerts, P.J. Bruggeman, M.
Cappelli, V. Colombo, U. Czarnetzki, U. Ebert, J.G. Eden, P.
Favia et al., J. Phys. D. Appl. Phys. 50, 323001 (2017)
65. D. Yan, Q. Wang, M. Adhikari, A. Malyavko, L. Lin, D.B.
Zolotukhin, X. Yao, M. Kirschner, J.H. Sherman, M. Keidar,
ACS Appl. Mater. Interfaces 12, 34548–34563 (2020)
66. N.K. Kaushik, N. Kaushik, K.C. Yoo, N. Uddin, J.S. Kim, S.J.
Lee, E.H. Choi, Biomaterials 87, 118–130 (2016)
67. D. Trachootham, J. Alexandre, P. Huang, Nat. Rev. Drug Dis-
cov. 8, 579–591 (2009)
68. J. Šimečková, F. Krčma, D. Klofáč, L. Dostál, Z. Kozáková,
Water 12, 2357 (2020)
69. S. Jung, H.J. Kim, S. Park, H.I. Yong, J.H. Choe, H.-J. Jeon,
W. Choe, C. Jo, Meat Sci. 108, 132–137 (2015)
70. P. Shaw, N. Kumar, H.S. Kwak, J.H. Park, H.S. Uhm, A.
Bogaerts, E.H. Choi, P. Attri, Sci. Rep. 8, 11268 (2018)
71. U. Schnabel, O. Handorf, K. Yarova, B. Zessin, S. Zechlin, D.
Sydow, E. Zellmer, J. Stachowiak, M. Andrasch, H. Below,
Foods 8, 55 (2019)
72. A. Azzariti, R.M. Iacobazzi, R. Di Fonte, L. Porcelli, R. Gris-
tina, P. Favia, F. Fracassi, I. Trizio, N. Silvestris, G. Guida
et al., Sci. Rep. 9, 1–13 (2019)
73. P. Attri, J.H. Park, A. Ali, E.H. Choi, Anti-Cancer Agents Med.
Chem. 18, 805–814 (2018)
74. N. Yoshikawa, W. Liu, K. Nakamura, K. Yoshida, Y. Ikeda, H.
Tanaka, M. Mizuno, S. Toyokuni, M. Hori, F. Kikkawa et al.,
Sci. Rep. 10, 1–8 (2020)
75. H.S. Uhm, E.H. Choi, G. Cho, Phys. Plasmas 7, 2744–2746
(2000)
76. H.S. Uhm, E.H. Choi, G.S. Cho, Appl. Phys. Lett. 78, 592–594
(2001)
77. M. Keidar, Plasma cancer therapy (Springer, 2020)
78. Evaluation guideline for plasma wound treatment (Ministry
of Food and Drug and Safety, Korean Government, 2016.12)”
(2016)
79. 21CFR801, “21CFR801.415 Maximum acceptable level of
ozone":"KS C9314 Standards of interior air cleaner (SPS-
KACA 002-0132)":"Criterion for chemical substance and
physical factor exposure" from Ministry of Employment and
Labor Notice #2020-48, Korea” (2020)
80. H.S. Uhm, E.H. Choi, G. Cho, K.W. Whang, Jpn. J. Appl. Phys.
40, L295 (2001)
81. G. Cho, Y.-G. Kim, Y.-S. Kim, D.-G. Joh, E.-H. Choi, Jpn. J.
Appl. Phys. 37, L1178 (1998)
82. H.D. Hagstrum, Phys. Rev. 96, 336 (1954)
83. J.Y. Lim, J.S. Oh, B.D. Ko, J. Won-Cho, S.O. Kang, G. Cho, H.S.
Uhm, E.H. Choi, J. Appl. Phys. 94, 764–769 (2003)
84. B. Ghimire, E.J. Szili, P. Lamichhane, R.D. Short, J.S. Lim, P.
Attri, K. Masur, K.-D. Weltmann, S.-H. Hong, E.H. Choi, Appl.
Phys. Lett. 114, 93701 (2019)
85. N. Srivastava, C. Wang, J. Appl. Phys. 110, 53304 (2011)
86. Y.J. Hong, C.J. Nam, K.B. Song, G.S. Cho, H.S. Uhm, D.I. Choi,
E.H. Choi, J. Instrum. 7, C03046 (2012)
87. B. Ghimire, J. Sornsakdanuphap, Y.J. Hong, H.S. Uhm, K.D.
Weltmann, E.H. Choi, Phys. Plasmas 24, 073502 (2017)
88. C. Bogdan, Nat. Immunol. 2, 907–916 (2001)
89. H.-P. Dorn, R. Neuroth, A. Hofzumahaus, J. Geophys. Res.
Atmos. 100, 7397–7409 (1995)
90. Y.H. Kim, Y.J. Hong, K.Y. Baik, G.C. Kwon, J.J. Choi, G.S. Cho,
H.S. Uhm, E.H. Choi et al., Plasma Chem. Plasma Process. 34,
457–472 (2014)
91. T.E.L. Smith, M.J. Wooster, M. Tattaris, D.W.T. Griffith, Atmos
Meas. Tech. 4, 97–116 (2011)
92. S. Kassi, K. Didriche, C. Lauzin, X. De Ghellinck D’Elseghem
Vaernewijckb, A. Rizopoulos, M. Herman, Spectrochim Acta
Part A Mol Biomol Spectrosc 75, 142–145 (2010)
93. Z. Du, S. Zhang, J. Li, N. Gao, K. Tong, Appl. Sci. 9, 338 (2019)
94. S. Iséni, S. Reuter, K.-D. Weltmann, J. Phys. D. Appl. Phys. 47,
75203 (2014)
95. J.P. Burrows, A. Dehn, B. Deters, S. Himmelmann, A. Rich-
ter, S. Voigt, J. Orphal, J. Quant. Spectrosc. Radiat. Transf. 60,
1025–1031 (1998)
96. Y.J. Hong, J. Lim, J.S. Choi, K.D. Weltmann, E.H. Choi, Plasma
Process. Polym. 18, 2000168 (2021)
97. X.-M. Zhu, Y.-K. Pu, J. Phys. D. Appl. Phys. 43, 403001 (2010)
98. X.-M. Zhu, Y.-K. Pu, Phys. Plasmas 12, 103501 (2005)
99. L.G. Piper, J. Chem. Phys. 97, 270–275 (1992)
100. V. Guerra, J. Loureiro, Plasma Sources Sci. Technol. 6, 361
(1997)
101. A. Lofthus, P.H. Krupenie, J. Phys. Chem. Ref. Data 6, 113–307
(1977)
102. S. Nunomura, M. Kondo, H. Akatsuka, Plasma Sources Sci.
Technol. 15, 783 (2006)
103. Y. Fuxiang, M. Zongxin, Z. Jialiang, Plasma Sci. Technol. 18, 79
(2016)
104. B. Ghimire, P. Lamichhane, J.S. Lim, B. Min, R. Paneru, K.-D.
Weltmann, E.H. Choi, Appl. Phys. Lett. 113, 194101 (2018)
105. B. Ghimire, E.J. Szili, P. Lamichhane, R.D. Short, J.S. Lim, P.
Attri, K. Masur, K.D. Weltmann, S.H. Hong, E.H. Choi, Appl.
Phys. Lett. 114, 093701 (2019)
106. P. Svarnas, Plasma Sci. Technol. 15, 891 (2013)
107. W. Tian, M.J. Kushner, J. Phys. D. Appl. Phys. 48, 494002 (2015)
108. A. Starikovskiy, Y. Yang, Y.I. Cho, A. Fridman, Plasma Sources
Sci. Technol. 20, 24003 (2011)
109. S. Kanazawa, H. Kawano, S. Watanabe, T. Furuki, S. Akamine,
R. Ichiki, T. Ohkubo, M. Kocik, J. Mizeraczyk, Plasma Sources
Sci. Technol. 20, 34010 (2011)
110. P. Attri, Y.H. Kim, D.H. Park, J.H. Park, Y.J. Hong, H.S. Uhm,
K.-N. Kim, A. Fridman, E.H. Choi, Sci. Rep. 5, 9332 (2015)
111. J.S. Lim, R.H. Kim, Y.J. Hong, P. Lamichhane, B.C. Adhikari,
J. Choi, E.H. Choi, Results Phys. 19, 103569 (2020)
112. J.S. Lim, Y.J. Hong, B. Ghimire, J. Choi, S. Mumtaz, E.H. Choi,
Results Phys. 20, 103693 (2021)
113. N.C. Roy, M.G. Hafez, M.R. Talukder, Phys. Plasmas 23, 83502
(2016)
114. P. Bruggeman, D.C. Schram, Plasma Sources Sci. Technol. 19,
045025 (2010)
115. D.-X. Liu, P. Bruggeman, F. Iza, M.-Z. Rong, M.G. Kong,
Plasma Sources Sci. Technol. 19, 25018 (2010)
116. Y. Itikawa, N. Mason, J. Phys. Chem. Ref. Data (2005). https://​
doi.​org/​10.​1063/1.​17992​51
13 Vol.:(0123456789)
850
117. IEC 60601-1-1. Medical electrical equipment—part 1–1: general
requirements for safety—colateral standard: safety requirements
for medical electrical systems
118. N. Leitgeb, Sicherheit von Medizingeräten: Recht-Risiko-Chan-
cen (Springer-Verlag, 2015)
119. M.S. Mann, R. Tiede, K. Gavenis, G. Daeschlein, R. Bussiahn,
K.-D. Weltmann, S. Emmert, T. von Woedtke, R. Ahmed, Clin.
Plasma Med. 4, 35–45 (2016)
120. A. Moldgy, G. Nayak, H.A. Aboubakr, S.M. Goyal, P.J. Brugge-
man, J. Phys. D. Appl. Phys. 53, 434004 (2020)
121. M.J. Nicol, T.R. Brubaker, B.J. Honish, A.N. Simmons, A.
Kazemi, M.A. Geissel, C.T. Whalen, C.A. Siedlecki, S.G. Bilén,
S.D. Knecht et al., Sci. Rep. 10, 1–11 (2020)
122. E.C. Filippidou, A. Koukouliata, Arch. Hell. Med. Ellenikes
Iatrikes, 28, (2011)
123. J. Gamble, W. Jones, S. Minshall, Environ. Res. 44, 6–17 (1987)
124. W.F. McDonnell, D.E. Abbey, N. Nishino, M.D. Lebowitz, Envi-
ron. Res. 80, 110–121 (1999)
125. J. Luo, A.F. Chen, Acta Pharmacol. Sin. 26, 259–264 (2005)
126. Y. Yang, P.K. Qi, Z.L. Yang, N. Huang, Biosurf. Biotribol. 1,
177–201 (2015)
127. P. Boukamp, R.T. Petrussevska, D. Breitkreutz, J. Hornung, A.
Markham, N.E. Fusenig, J. Cell Biol. 106, 761–771 (1988)
128. R. Bussiahn, N. Lembke, R. Gesche, T. von Woedtke, K.D. Welt-
mann, Hyg. Med 38, 212–216 (2013)
129. B.R.W.K.-D.R.M.K.R.P.F. Metelmann H-R von Woedtke Th,
W.P. D, Am. J. Cosmet. Surg., 29, 52 (2012)
130. P. Attri, K. Koga, M. Shiratani, Appl. Phys. Express 14, 27002
(2021)
131. M.-S. Choi, E.B. Jeon, J.Y. Kim, E.H. Choi, J.S. Lim, J. Choi,
S.Y. Park, J. Food Eng. 278, 109952 (2020)
132. M. Adhikari, N. Kaushik, B. Ghimire, B. Adhikari, S. Baboota,
A.A. Al-Khedhairy, R. Wahab, S.-J. Lee, N.K. Kaushik, E.H.
Choi, Cell Commun Signal. 17, 1–14 (2019)
133. A. Nasir, G. Caetano-Anollés, Sci. Adv. 1, e1500527 (2015)
134. G. Bauer, Plasma Med. 9, 1 (2019)
135. G. Bauer, Redox Biol. 26, 101291 (2019)
136. G. Bauer, D. Sersenová, D.B. Graves, Z. Machala, Sci. Rep. 9,
1–28 (2019)
137. A. Dubuc, P. Monsarrat, F. Virard, N. Merbahi, J.P. Sarrette,
S. Laurencin-Dalicieux, S. Cousty, Therap Adv Med Oncol 10,
1–12 (2018)
138. X. Zhou, D. Cai, S. Xiao, M. Ning, R. Zhou, S. Zhang, X. Chen,
K. Ostrikov, X. Dai, J. Cancer 11, 2273 (2020)
139. F. Saadati, H. Mahdikia, H.-A. Abbaszadeh, M.-A. Abdollahifar,
M.S. Khoramgah, B. Shokri, Sci. Rep. 8, 1–15 (2018)
140. L. Xiang, X. Xu, S. Zhang, D. Cai, X. Dai, Free Radic. Biol.
Med. 124, 205–213 (2018)
141. X. Cheng, W. Rowe, L. Ly, A. Shashurin, T. Zhuang, S. Wigh, G.
Basadonna, B. Trink, M. Keidar, J. Canady, Plasma 1, 218–228
(2018)
142. M. Keidar, D. Yan, I.I. Beilis, B. Trink, J.H. Sherman, Trends
Biotechnol. 36, 586–593 (2018)
143. K.J. Schlegel, B. Veronika, Clin. Plasma Med. 1, 2 (2013)
144. D. Yan, H. Cui, W. Zhu, N. Nourmohammadi, J. Milberg, L.G.
Zhang, J.H. Sherman, M. Keidar, Sci. Rep. 7, 1–12 (2017)
145. G.I. Deichman, Biochemistry 65, 78–94 (2000)
146. B. Böhm, S. Heinzelmann, M. Motz, G. Bauer, Biol. Chem. 396,
1339–1356 (2015)
147. G. Bauer, D.B. Graves, Plasma Process. Polym. 13, 1157–1178
(2016)
148. H. Mokhtari, L. Farahmand, K. Yaserian, N. Jalili, K. Majidza-
deh-A, J. Cell. Physiol. 234, 6778–6782 (2019)
149. A. Lin, Y. Gorbanev, J. De Backer, J. Van Loenhout, W. Van
Boxem, F. Lemière, P. Cos, S. Dewilde, E. Smits, A. Bogaerts,
Adv. Sci. 6, 1802062 (2019)
13 Vol.:(0123456789)
E. H. Choi et al.
150. A. Lin, B. Truong, S. Patel, N. Kaushik, E.H. Choi, G. Frid-
man, A. Fridman, V. Miller, Int. J. Mol. Sci. 18, 966 (2017)
151. A.G. Lin, B. Xiang, D.J. Merlino, T.R. Baybutt, J. Sahu,
A. Fridman, A.E. Snook, V. Miller, Oncoimmunology 7,
e1484978 (2018)
152. S. Bekeschus, J. Kolata, C. Winterbourn, A. Kramer, R. Turner,
K.D. Weltmann, B. Bröker, K. Masur, Free Radic. Res. 48,
542–549 (2014)
153. H.J. Ahn, K. Il-Kim, N.N. Hoan, C.H. Kim, E. Moon, K.S.
Choi, S.S. Yang, J.-S. Lee, PLoS ONE 9, e86173 (2014)
154. N.K. Kaushik, N. Kaushik, B. Min, K.H. Choi, Y.J. Hong, V.
Miller, A. Fridman, E.H. Choi, J. Phys. D. Appl. Phys. 49,
84001 (2016)
155. N.K. Kaushik, N. Kaushik, M. Adhikari, B. Ghimire, N.N.
Linh, Y.K. Mishra, S.-J. Lee, E.H. Choi, Cancers (Basel) 11,
842 (2019)
156. F. Virard, S. Cousty, J.-P. Cambus, A. Valentin, P. Kémoun, F.
Clément, PLoS ONE 10, e0133120 (2015)
157. C. Schneider, L. Gebhardt, S. Arndt, S. Karrer, J.L. Zimmer-
mann, M.J.M. Fischer, A.-K. Bosserhoff, Sci. Rep. 8, 1–10
(2018)
158. K. Wende, S. Straßenburg, B. Haertel, M. Harms, S. Holtz, A.
Barton, K. Masur, T. von Woedtke, U. Lindequist, Cell Biol.
Int. 38, 412–425 (2014)
159. A.M. Hirst, M.S. Simms, V.M. Mann, N.J. Maitland, D.
O’connell, F.M. Frame, Br. J. Cancer 112, 1536–1545 (2015)
160. J. Köritzer, V. Boxhammer, A. Schäfer, T. Shimizu, T.G.
Klämpfl, Y.-F. Li, C. Welz, S. Schwenk-Zieger, G.E. Morfill,
J.L. Zimmermann, PLoS ONE 8, e64498 (2013)
161. S. Lee, H. Lee, D. Jeong, J. Ham, S. Park, E.H. Choi, S.J. Kim,
Free Radic. Biol. Med. 110, 280–290 (2017)
162. I. Eke, N. Cordes, Semin. Cancer Biol. 31, 65–75 (2015)
163. N.K. Kaushik, N. Kaushik, N.N. Linh, B. Ghimire, A. Pengkit,
J. Sornsakdanuphap, S.J. Lee, E.H. Choi, Nanomaterials 9,
1–19 (2019)
164. L.N. Nguyen, N. Kaushik, P. Lamichhane, S. Mumtaz, R.
Paneru, P. Bhartiya, J.S. Kwon, Y.K. Mishra, L.Q. Nguyen,
N.K. Kaushik et al., Green Chem. 22, 6588–6599 (2020)
165. X. Dai, K. Bazaka, D.J. Richard, E.R.W. Thompson, K.K.
Ostrikov, Trends Biotechnol. 36, 1183–1198 (2018)
166. E. Biscop, A. Lin, W. Van Boxem, J. Van Loenhout, J. De
Backer, C. Deben, S. Dewilde, E. Smits, A. Bogaerts, Cancers
(Basel) 11, 1287 (2019)
167. W. Van Boxem, J. der Paal, Y. Gorbanev, S. Vanuytsel, E.
Smits, S. Dewilde, A. Bogaerts, Sci. Rep. 7, 1–15 (2017)
168. P.-M. Girard, A. Arbabian, M. Fleury, G. Bauville, V. Puech,
M. Dutreix, J.S. Sousa, Sci. Rep. 6, 1–17 (2016)
169. N. Kurake, H. Tanaka, K. Ishikawa, T. Kondo, M. Sekine, K.
Nakamura, H. Kajiyama, F. Kikkawa, M. Mizuno, M. Hori,
Arch. Biochem. Biophys. 605, 102–108 (2016)
170. E. Griseti, N. Merbahi, M. Golzio, Cancers (Basel) 12, 721
(2020)
171. K.R. Liedtke, S. Bekeschus, A. Kaeding, C. Hackbarth, J.-P.
Kuehn, C.-D. Heidecke, W. von Bernstorff, T. von Woedtke,
L.I. Partecke, Sci. Rep. 7, 1–12 (2017)
172. G. Isbary, J. Köritzer, A. Mitra, Y.-F. Li, T. Shimizu, J.
Schroeder, J. Schlegel, G.E. Morfill, W. Stolz, J.L. Zimmer-
mann, Clin. Plasma Med. 1, 36–44 (2013)
173. J. Gay-Mimbrera, M.C. Garcia, B. Isla-Tejera, A. Rodero-
Serrano, A.V. Garcia-Nieto, J. Ruano, Adv. Ther. 33, 894–909
(2016)
174. Z. Xiong, Trends Biotechnol. 36, 582–583 (2018)
175. M. Akter, A. Jangra, S.A. Choi, E.H. Choi, I. Han, Cancers
(Basel) 12, 245 (2020)
176. M. Akter, J.S. Lim, E.H. Choi, I. Han, Cells 10, 236 (2021)
Plasma bioscience for medicine, agriculture and hygiene applications 851
177. Y. Li, M.H. Kang, H.S. Uhm, G.J. Lee, E.H. Choi, I. Han, Sci.
Rep. 7, 1–9 (2017)
178. C.-H. Song, P. Attri, S.-K. Ku, I. Han, A. Bogaerts, E.H. Choi,
J. Phys. D. Appl. Phys. 54, 185202 (2021)
179. Y. Li, J.H. Kim, E.H. Choi, I. Han, Sci. Rep. 9, 1–10 (2019)
180. I. Han, E.H. Choi, Oncotarget 8, 36399 (2017)
181. Y. Li, E.H. Choi, I. Han, Oxid. Med. Cell. Longev. 2019,
2318680 (2019)
182. S.W. Wilhelm, C.A. Suttle, Bioscience 49, 781–788 (1999)
183. F.G. Dobrynin, D. Fridman, F.A. Fridman, New J. Phys. 11,
115020 (2009)
184. C. Tendero, C. Tixier, P. Tristant, J. Desmaison, P. Leprince,
Spectrochim. Acta Part B At. Spectrosc. 61(2), 30 (2006)
185. M. Laroussi, IEEE Trans. Plasma Sci. 24, 1188–1191 (1996)
186. S. Bekeschus, A. Kramer, E. Suffredini, T. Von Woedtke, V.
Colombo, IEEE Trans. Radiat. Plasma Med. Sci. 4, 391–399
(2020)
187. Z. Chen, G. Garcia Jr., V. Arumugaswami, R.E. Wirz, Phys. Flu-
ids 32, 111702 (2020)
188. M. Weiss, G. Daeschlein, A. Kramer, M. Burchardt, S. Brucker,
D. Wallwiener, M.B. Stope, J. Med. Virol. 89, 952–959 (2017)
189. L.N. Nguyen, N. Kaushik, P. Bhartiya, S.K. Gurmessa, H.-J.
Kim, L.Q. Nguye, N.K. Kaushik, E.H. Choi, J. Ind. Eng. Chem.
(2021). https://​doi.​org/​10.​1016/j.​jiec.​2021.​05.​035
190. P. Attri, N.K. Kaushik, N. Kaushik, D. Hammerschmid, A.
Privat-Maldonado, J. De Backer, M. Shiratani, E.H. Choi, A.
Bogaerts, Int. J. Biol. Macromol. 182, 1724 (2021)
191. A. Barjasteh, Z. Dehghani, P. Lamichhane, N. Kaushik, E.H.
Choi, N.K. Kaushik, Appl. Sci. 11, 3372 (2021)
192. N.K. Kaushik, S. Bekeschus, H. Tanaka, A. Lin, E.H. Choi, Appl.
Sci. 11, 4584 (2021)
193. P. Lamichhane, M. Veerana, J.S. Lim, S. Mumtaz, B. Shrestha,
N.K. Kaushik, G. Park, E.H. Choi, Int. J. Mol. Sci. 22, 5360
(2021)
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